Methanogenic and sulphate reducing bacterial population levels in a full-scale anaerobic reactor treating pulp and paper industry wastewater using fluorescence in situ hybridisation.

In this study, specific methanogenic activity (SMA) test and fluorescence in situ hybridisation (FISH) were respectively used to determine acetoclastic methanogenic capacity, and composition and number of methanogenic and sulphate reducing bacterial (SRB) populations within a full scale anaerobic contact reactor treating a pulp and paper industry effluent. The sludge samples were collected from three different heights along the anaerobic reactor having a difficulty of completely stirring. Performance of the anaerobic reactor in terms of COD removal efficiency varied between 47 and 55% at organic loading rates in a range of 1.6-1.8 kg COD m(-3) d(-1) and methane yield varied between 0.18 and 0.20 m3CH4kg CODrem(-1). The anaerobic reactor was not operated for 2 weeks during the monitoring period. According to SMA test results, potential methane production rate was 276 mLCH4 gVSS(-1) d(-1) before the off period of the reactor, however it decreased to 159 mL CH4 gVSS(-1) d(-1) after this period. SMA test and FISH results along the reactor height showed that the acetoclastic methanogenic activity of the sludge samples, the relative abundance of acetoclastic methanogens, hydrogenotrophic methanogens and acetate oxidising SRB decreased as the reactor height increased, however the relative abundance of non-acetate oxidising SRB increased.     

On-site treatment of high-strength alcohol distillery wastewater by a pilot-scale thermophilic multi-staged UASB (MS-UASB) reactor.

A pilot-scale multi-staged UASB (MS-UASB) reactor with a working volume of 2.5 m3 was operated for thermophilic (55 degrees C) treatment of an alcohol distillery wastewater for a period of over 600 days. The reactor steadily achieved a super-high rate COD removal, that is, 60 kgCOD m(-3) d(-1) with over 80% COD removal. However, when higher organic loading rates were further imposed upon the reactor, that is, above 90 kgCOD m(-3) d(-1) for barely-based alcohol distillery wastewater (ADWW) and above 100 kgCOD m(-3) d(-1) for sweet potato-based ADWW, the reactor performance somewhat deteriorated to 60 and 70% COD removal, respectively. Methanogenic activity (MA) of the retained sludge in the thermophilic MS-UASB reactor was assessed along the time course of continuous run by serum-vial test using different substrates as a vial sole substrate. With the elapsed time of continuous run, hydrogen-utilizing MA, acetate-utilizing MA and propionate-fed MA increased at maximum of 13.2, 1.95 and 0.263 kgCOD kgVSS(-1) d(-1), respectively, indicating that propionate-fed MA attained only 1/50 of hydrogen-utilizing MA and 1/7 of acetate-utilizing MA. Since the ADWW applied herewith is a typical seasonal campaign wastewater, the influence of shut-down upon the decline in sludge MA was also investigated. Hydrogen-utilizing MA and acetate-utilizing MA decreased slightly by 3/4, during a month of non-feeding period, whereas propionate-fed MA were decreased significantly by 1/5. Relatively low values of propionate-fed MA and its vulnerability to adverse conditions suggests that the propionate degradation step is the most critical bottleneck of overall anaerobic degradation of organic matters under thermophilic condition.     

Full scale fluidized bed anaerobic reactor for domestic wastewater treatment: performance, sludge production and biofilm.

This paper describes the performance, sludge production and biofilm characteristics of a full scale fluidized bed anaerobic reactor (32 m3) for domestic wastewater treatment. The reactor was operated with 10.5 m x h(-1) upflow velocity, 3.2 h hydraulic retention time, and recirculation ratio of 0.85 and it presented removal efficiencies of 71+/-8% of COD and 77+/-14% of TSS. During the apparent steady-state period, specific sludge production and sludge age in the reactor were (0.116+/-0.033) kgVSS. kgCOD(-1) and (12+/-5)d, respectively. Biofilm formed in the reactor presented two different patterns: one of them at the beginning of the colonization and the other of mature biofilm. These different colonization patterns are due to bed stratification in the reactor, caused by the difference in local-energy dissipation rates along the reactor's height, and density, shape, etc. of the bioparticles. The biofilm population is formed mainly of syntrophic consortia among sulfate reducing bacteria, methanogenic archaea such as Methanobacterium and Methanosaeta-like cells.     

Start-up and steady-state results of a full-scale UASB reactor treating malting wastewater.

An Imhoff tank was reconstructed into a 250 m3 UASB reactor in order to treat a malting plant wastewater. The UASB was inoculated with sludge from an anaerobic lagoon used for slaughterhouse wastewater treatment. After two months of operation the reactor achieved full load with an HRT of 17 h, a COD removal higher than 80% and a biogas production of 300 m3/day (77% average methane content), with an organic loading rate of 3.6 kgCOD/m3.d (0.24 kgCOD/kgVSS.d). A yield coefficient of 0.09 gVSS/gCODrem was found from a mass balance. The fat present in the inoculated sludge (48 mg/gSSV) did not affect the start up performance. Sludge from the inoculum with high content of fat (270 mg/gSSV), was separated by flotation in the first week of operation. The COD removal efficiency was scarcely influenced by the reactor operation temperature (17-25 degrees C).     

Anaerobic co-digestion of winery wastewater.

The operational performance of anaerobic batch reactors treating winery wastewater (WW) combined with waste activated sludge (WAS) in different proportions was investigated under mesophilic conditions. In these experiments it was shown that for anaerobic digestion of WW alone, methane production rate was lower than the rates achieved when WW and WAS were treated together. When WW was mixed with WAS at a concentration of 50% WW resulted in the highest methane production rates. A simplified anaerobic model was used to determine the main kinetic parameters; maximum COD reduction rate (q(DA)) and maximum methane generation rate (k(max)). The maximum values of q(DA) and k(max) were 16.50 kgCOD COD(-1) d(-1) and 14.34 kgCOD kgCOD(-1) d(-1), respectively.     

Sludge deterioration in a full scale UASB reactor after a pH drop working under low loading conditions.

A full scale UASB reactor treating the effluent of a malting plant was operated during nearly two years. During 37 weeks of operation the reactor worked with a COD removal efficiency of 80% and a biogas production of nearly 300 m(3)/d with a methane content of 77%. After the start up and during these months of operation the volumetric organic load was 4 kgCOD/m(3).d and the specific organic load was between 0.2-0.4 kgCOD/kgVSS.d. The sludge SMA in this period was around 0.25 kgCOD/kg VSS.d. On week 37 as a result of a problem at the industrial process the pH in the reactor dropped to a value of 4.8. After pH recovering, the reactor worked with fluctuating COD values in the exit and showed a downward trend in the COD removal efficiency. On week 81 the presence of filaments in the granules was detected. High proportion of Chloroflexi filaments were detected by FISH in the sludge. Changes in the microbial population caused by the low pH probably destabilize the reactor performance. The presence of filamentous granules in the sludge and its further growing could be encouraged by the pH drop and the low specific organic load applied to the reactor. The low specific organic load was a consequence of the high VSS content in the UASB reactor, due to the lack of purges. The length of the filaments attached to the granules grew throughout time. In order to eliminate the sludge with poor settlement properties a recycle was applied to the reactor. As a consequence, low amount of granular sludge stayed in the reactor. At the end, COD concentration in the influent reached higher values than in normal operation; at the same time a complete sludge wash out occurred. On the other hand, using the same sludge (after the recycle implementation) in a bench scale reactor the good properties of the sludge were completely recovered.     

High rate anaerobic thermophilic technologies for distillery wastewater treatment.

In this paper, performance of two high rate technologies, upflow anaerobic fixed-film reactor and fluidized bed laboratory-scale, treating distillery wastewater (wine vinasses) at anaerobic thermophilic conditions have been compared. The results obtained show that the stationary packed bed, with a corrugated plastic support, operated under stable conditions at organic loading rates (OLR0) around 20 kgCOD/m3/d, gives maximal total CODr of 76% at OLR0 of 6.29 kgCOD/m3/d; the fluidized bed reactor, operated on open pore sintered-glass media, gives total CODr of 96% at OLR0 of 5.88 kgCOD/m3/d. The anaerobic fluidized bed technology is more effective than the upflow anaerobic fixed-film technology due, fundamentally, to this technology favouring the transport of microbial cells from the bulk to the surface and enhancing the contact between the microorganism-substrate phases, In this sense, the stationary packed bed technology is adequate for the treatment of easily biodegradable wastewater, or for the cases where elevated percentages of CODr removal are not required, while the fluidized bed technology is especially suitable for treatment of hazardous wastes with recalcitrant compositions.     

Pretreatment of silk-dyeing industrial wastewater by UASB reactor.

The objective of this study was to investigate the performance of the upflow anaerobic sludge blanket (UASB) reactor as the pretreatment system for silk-dyeing wastewater. Two laboratory-scale UASB reactors, with working volume of 15.59 I, were used during May 1998 to June 1999. The actual wastewater was diluted to reduce ammonium ion toxicity on anaerobic bacteria. The experiments were conducted at the organic loading rates (OLRs) of 0.52, 1.01, 1.04, 1.54 and 2.56 kgCOD/(m3 x d), treating only wastewater generated from the acid-dye process of mixed-species raw silk. It took approximately 4 1/2 months to reach the steady-state conditions. It was found that the COD removal was in the ranges of 74.1-85.3%, except at OLR 2.56 kgCOD/(m3 x d) where efficiency significantly dropped to 55.2%. The apparent color removal was in the similar trend as COD. During the study periods, wastewater input had various color shades while the effluent generally looked pale yellowish. The methane generation rates ranged from 0.18-0.31 m3/kg COD removed, with methane composition 81.0-88.1% in biogas. The average granule size in the sludge bed had slowly increased to 0.73 mm in the last experiment. It can be concluded that the UASB reactor is suitable as a pretreatment system for silk-dyeing wastewater. An OLR of 1 kgCOD/(m3 x d) and an influent concentration diluted to 2,600 mgCOD/l are suggested while COD and apparent color removal efficiency of 80% and 70%, respectively, can be expected.     

Thermal pre-treatment of primary and secondary sludge at 70 degrees C prior to anaerobic digestion.

In general, mesophilic anaerobic digestion of sewage sludge is more widely used compared to thermophilic digestion, mainly because of the lower energy requirements and higher stability of the process. However, the thermophilic anaerobic digestion process is usually characterised by accelerated biochemical reactions and higher growth rate of microorganisms resulting in an increased methanogenic potential at lower hydraulic retention times. Furthermore, thermal pre-treatment is suitable for the improvement of stabilization and could be realized at relatively low cost especially at low temperatures. The present study investigates the effect of the pre-treatment at 70 degrees C on thermophilic (55 degrees C) anaerobic digestion of primary and secondary sludge in continuously operated digesters. Thermal pre-treatment of primary and secondary sludge at 70 degrees C enhanced the removal of organic matter and the methane production during the subsequent anaerobic digestion step at 55 degrees C. It also greatly contributed to the destruction of pathogens present in primary sludge. Finally it results in enhanced microbial activities of the subsequent anaerobic step suggesting that the same efficiencies in organic matter removal and methane recovery could be obtained at lower HRTs.     

Effect of thermal alkaline pretreatment on the anaerobic digestion of wasted activated sludge

The effect of alkaline pretreatment of waste-activated sludge, using two models to study the sequential hydrolysis rates of suspended (Sanders' surface model) and dissolved (Goel's saturation model) solids, on the mesophilic and thermophilic anaerobic digestion rate is evaluated. The pretreatment, which reduces the size of the solids, increases the reaction rate by increasing the surface area and the specific surface hydrolysis constant (K(SBK)); at thermophilic conditions from 0.45 x 10(-3) kg m(-2) d(-1) for the fresh sludge to 0.74 x 10(-3) kg m(-2) d(-1) for the pretreated sludge and at mesophilic conditions these values are 0.28 x 10(-3) kg m(-2) d(-1) and 0.47 x 10(-3) kg m(-2) d(-1) confirming the usefulness of a pretreatment for solids reduction. But for soluble solids, the thermoalkaline pretreatment decreases the reaction rates by inducing a competitive inhibition on the thermophilic anaerobic digestion rate while in the mesophilic range, a non-competitive inhibition is observed. A mathematical simulation of the consecutive reactions, suspended solids to dissolved solids and to methane in staged anaerobic thermophilic-mesophilic digestion, shows that with 4% suspended solids concentration it is better not to use a thermoalkaline pretreatment because overall solids reduction and total methane production are not as good as without pretreatment.     

Effect of the pH influent conditions in fixed-film reactors for anaerobic thermophilic treatment of wine-distillery wastewater.

In anaerobic treatments, the pH conditions affect the efficacy and operation of the process. The main purpose of this research is to compare the effect of the pH influent on the performance of a high rate technology at laboratory scale, upflow anaerobic fixed-film reactor, treating distillery wastewater (wine vinasses) in thermophilic conditions. The results obtained shown that the pH influent influences the performance of the biodegradation process: the depurative efficiency is higher for the operation with alkaline influent. The operation with acid influent allows us to operate at organic loading rates (OLR) around 5.6 kgCOD/m3/d (hydraulic retention time: 1.5 days), maintaining total Chemical Oxygen Demand removals (CODr) of 77.2%; the operation with alkaline influent allows total CODr of 76.8% working at OLR around 10.5 kgCOD/m3/d. The greatest efficiency of substrate removal was 87.5% for OLR 3.2 kgCOD/m3/d and hydraulic retention time of 4.0 days operating with alkaline influent. Therefore, the operation with alkaline influent implicates senior levels of purifying efficiency for similar organic load rate.     

Maize mono-digestion efficiency: results from laboratory tests

A laboratory experimental campaign was carried out in order to assess the optimal configuration for the anaerobic digestion of a mixture of sweet corn and ensiled maize. Batch hydrolysis tests were conducted at 35 and 55 °C and at four different particle sizes (2, 5, 20 and 50 mm) obtained by manual chopping and sieving. Chemical pre-treatment by 24 h incubation at various acid and alkaline pH was also considered for its potential to increase the maize methane yield. Results suggest that the hydrolytic phase proceeds significantly faster under thermophilic conditions. Significant differences in the solubilization rate were also observed when comparing coarse (20-50 mm) with fine (2-5 mm) particles, while 2 and 5 mm particles were solubilized at similar rates. No advantages from the chemical pre-treatment, in terms of solubilization efficiency and biomethanization potential were observed. According to these preliminary results, a two-stage semi-continuous laboratory plant consisting of a thermophilic hydrolytic reactor followed by a mesophilic methanogenic reactor was operated for 110 days. Steady state loading parameters were: influent concentration (maize mixture diluted in tap water) of 46 g VS/L, hydraulic retention time of 31 d, organic loading rate of 1.5 g VS/L/d. Alkalinity was dosed to the methanogenic reactor to avoid pH drops. Collected data allowed the average biodegradation efficiency to be estimated at around 60-65%.     

Evaluation of sludge properties in a pilot-scale UASB reactor for sewage treatment in a temperate region

In this study, continuous operation of a pilot-scale upflow anaerobic sludge blanket (UASB) reactor for sewage treatment was conducted for 630 days to investigate the physical and microbial characteristics of the retained sludge. The UASB reactor with a working volume of 20.2 m(3) was operated at ambient temperature (16-29 °C) and seeded with digested sludge. After 180 days of operation, when the sewage temperature had dropped to 20 °C or lower, the removal efficiency of both total suspended solids (TSS) and total biochemical oxygen demand (BOD) deteriorated due to washout of retained sludge. At low temperature, the cellulose concentration of the UASB sludge increased owing to the rate limitation of the hydrolytic reaction of suspended solids in the sewage. However, after an improvement in sludge retention (settleability and concentration) in the UASB reactor, the process performance stabilized and gave sufficient results (68% of TSS removal, 75% of total BOD removal) at an hydraulic retention time (HRT) of 9.7 h. The methanogenic activity of the retained sludge significantly increased after day 246 due to the accumulation of Methanosaeta and Methanobacterium following the improvement in sludge retention in the UASB reactor. Acid-forming bacteria from phylum Bacteroidetes were detected at high frequency; thus, these bacteria may have an important role in suspended solids degradation.     

Anaerobic thermophilic digestion of sewage sludge with a thickened sludge recycle

The process of anaerobic thermophilic digestion of municipal wastewater sludge with a recycled part of thickened digested sludge, was studied in semi-continuous laboratory digesters. This modified recycling process resulted in increased solids retention time (SRT) with the same hydraulic retention time (HRT) as compared with traditional digestion without recycling. Increased SRT without increasing of HRT resulted in the enhancement of volatile substance reduction by up to 68% in the reactor with the recycling process compared with 34% in a control conventional reactor. Biogas production was intensified from 0.3 L/g of influent volatile solids (VS) in the control reactor up to 0.35 L/g VS. In addition, the recycling process improved the dewatering properties of digested sludge.     

Relative kinetics of anaerobic digestion under thermophilic and mesophilic conditions

With several advantages over the conventional mesophilic anaerobic digestion, such as better sludge quality and higher biogas production, thermophilic anaerobic digestion is regarded as a promising alternative for sludge digestion. Primary and activated sludges are complex materials, and historically, analysis of kinetics has been largely on whole sludge, without analysis of individual components. This paper analyses relative digestion kinetics of pure substrates designed to target main stages of sludge digestion under thermophilic and mesophilic conditions. Hydrolysis rate of cellulose was significantly influenced by temperature with hydrolysis coefficients of--at 55 degrees C (0.7 +/- 0.1 day(-1)), 60 degrees C (0.8 +/- 0.2 day(-1)), 65 degrees C (1.1 +/- 0.2 day(-1)) and 70 degrees C (1.2 +/- 0.2 day(-1)) over 38 degrees C (0.4 +/- 0.1 day(-1)). This strongly follows the Arrhenius relationship, with an activation energy (E(A)) of 31 +/- 4 kJ mol(-1), corresponding to an increase of 1.5x for each 10 degrees C of temperature increase. Glucose uptake was rapid with a wide variety of fermentation products detected under mesophilic conditions, while uptake was slower under thermophilic conditions with acetate and propionate being dominant products. Propionate acetogenesis and acetate-utilizing methanogenesis kinetics were not influenced by temperatures. Hydrolysis is widely regarded as a rate-limiting step in sludge digestion, thus improvements in hydrolysis rates as measured during this study have the potential for significant improvements in overall apparent sludge digestion rates.     

Real-time quantifications of dominant anaerobes in an upflow reactor by polymerase chain reaction using a TaqMan probe.

This study was conducted to demonstrate the application of quantitative real-time polymerase chain reaction (qRT-PCR) for the quantification of dominant bacteria in an anaerobic reactor using a designed TaqMan probe. A novel group of uncultured thermophilic bacteria affiliated with Thermotogales was first found in a phenol-degrading sludge from a 55 degrees C upflow anaerobic sludge blanket (UASB) reactor, which effectively removed 99% of phenol at loading of 0.51 g-phenol l(-1) d(-1) h of hydraulic retention. A TaqMan probe was then designed targeting this group of Thermotogales affiliated bacteria (TAB), and used to monitor its concentration in the reactors. Results showed that the TAB population in the 55 degrees C reactor increased proportional to the phenol degrading rate. Results also showed that the TAB population ranged 3.5-9.9% in the 55 degrees C phenol-degrading sludge, but only 0.0044% in the 37 degrees C sludge and 0.000086% in the 26 degrees C sludge.     

Microbial characteristics of a methanogenic phenol-degrading sludge.

Microbial properties of a methanogenic granular phenol-degrading sludge were characterized using the 16S rRNA/DNA-based techniques, including polymerase chain reaction (PCR) amplification, cloning, DNA sequencing, and fluorescence in situ hybridization (FISH). The sludge was sampled from an upflow anaerobic sludge blanket reactor, which removed 98% of phenol (up to 1260 mg/l) in wastewater at 26 degrees C with 12 hours of hydraulic retention. Based on DNA analysis, the Eubacteria in the sludge was composed of 13 operational taxonomy units (OTUs). Two OTUs, one resembling Clostridium and the other remotely resembling Desulfotomaculum, were likely responsible for the conversion of phenol to benzoate, which was further degraded by five Syntrophus-resembling OTUs to acetate and H2/CO2; methanogens lastly converted acetate and H2/CO2 into methane. The role of six remaining OTUs remains unclear. Overall, the sludge was composed of 26 +/- 6% Eubacteria and 74 +/- 9% methanogens, of which 54 +/- 6% were acetotrophic Methanosaetaceae, 14 +/- 3% and 3 +/- 2% were hydrogenotrophic Methanomicrobiales and Methanobacteriaceae, respectively.     

Thermophilic anaerobic digestion in compact systems: investigations by modern microbiological techniques and mathematical simulation.

Thermophilic anaerobic digestion in compact systems can be an economical and ecological reasonable decentralised process technique, especially for rural areas. Thermophilic process conditions are important for a sufficient removal of pathogens. The high energy demand, however, can make such systems unfavourable in terms of energy costs. This is the case when low concentrated wastewater is treated or the system is operated at low ambient temperatures. In this paper we present experimental results of a compact thermophilic anaerobic system obtained with fluorescent in situ hybridisation (FISH) analysis and mathematical simulation. The system was operated with faecal sludge for a period of 135 days and with a model substrate consisting of forage and cellulose for a period of 60 days. The change in the microbial community due to the two different substrates treated could be well observed by the FISH analysis. The Anaerobic Digestion Model no. 1 (ADM1) was used to evaluate system performance at different temperature conditions. The model was extended to contribute to decreased methanogenic activity at lower temperatures and was used to calculate energy production. A model was developed to calculate the major parts of energy consumed by the digester itself at different temperature conditions. It was demonstrated by the simulation study that a reduction of the process temperature can lead to higher net energy yield. The simulation study additionally showed that the effect of temperature on the energy yield is higher when a substrate is treated with high protein content.     

Autogenerative high pressure digestion: anaerobic digestion and biogas upgrading in a single step reactor system

Conventional anaerobic digestion is a widely applied technology to produce biogas from organic wastes and residues. The biogas calorific value depends on the CH, content which generally ranges between 55 and 65%. Biogas upgrading to so-called 'green gas', with natural gas quality, generally proceeds with add-on technologies, applicable only for biogas flows > 100 m3/h. In the concept of autogenerative high pressure digestion (AHPD), methanogenic biomass builds up pressure inside the reactor. Since CO2 has a higher solubility than CH4, it will proportion more to the liquid phase at higher pressures. Therefore, AHPD biogas is characterised by a high CH4 content, reaching equilibrium values between 90 and 95% at a pressure of 3-90 bar. In addition, also H2S and NH3 are theoretically more soluble in the bulk liquid than CO2. Moreover, the water content of the already compressed biogas is calculated to have a dew point <--10 degrees C. Ideally, high-quality biogas can be directly used for electricity and heat generation, or injected in a local natural gas distribution net. In the present study, using sodium acetate as substrate and anaerobic granular sludge as inoculum, batch-fed reactors showed a pressure increase up to 90 bars, the maximum allowable value for our used reactors. However, the specific methanogenic activity (SMA) of the sludge decreased on average by 30% compared to digestion at ambient pressure (1 bar). Other results show no effect of pressure exposure on the SMA assessed under atmospheric conditions. These first results show that the proposed AHPD process is a highly promising technology for anaerobic digestion and biogas upgrading in a single step reactor system.     

Mesophilic-thermophilic-mesophilic anaerobic digestion of liquid dairy cattle manure.

The potential of a mesophilic-thermophilic-mesophilic anaerobic digestion system was investigated with respect to improvement of both digestion and sanitation efficiencies during treatment of liquid cattle manure. The pilot plant produced a high methane yield from liquid dairy cattle manure of 0.24 m3 (kg VSfed)(-1) Considering the low system loading rate of 1.4-1.5 kg VS (m3 d)(-1), digestion efficiency compared to conventional processes did not appear improved. The minimum guaranteed retention time in the tubular thermophilic reactor was increased compared to a continuously stirred tank reactor. Levels of intestinal enterococci in raw liquid manure as determined with cultivation methods were reduced by 2.5 -3 log units to a level of around 10(2) cfu/mL. This sanitizing effect was achieved both during mesophilic-thermophilic-mesophilic and thermophilic-mesophilic treatment, provided the thermophilic digester was operated at 53-55 degrees C. A change in feeding interval from 1 h to 4 h did not significantly alter methane yield and sanitation efficiency. It was proposed that a two-stage, thermophilic-mesophilic anaerobic digestion system would be able to achieve the same sanitizing effect and equal or better digestion efficiency at lower costs.