Thermophilic Sludge Research Articles

Digestion Performance of Various Combinations of Thermophilic and Mesophilic Sludge Digestion Systems

Digestion Performance of Various Combinations of Thermophilic and Mesophilic Sludge Digestion Systems

Sludge properties and their effects on membrane fouling in submerged anaerobic membrane bioreactors (SAnMBRs)

Two submerged anaerobic membrane bioreactors (SAnMBRs) (thermophilic vs. mesophilic) were operated for a period of 3.5 months with kraft evaporator condensate at a feed chemical oxygen demand of 10,000 mg/L. The results show that the filtration behavior of the two systems was significantly different. The filtration resistance in the thermophilic SAnMBR was about 5–10 times higher than that of the mesophilic system when operated under similar hydrodynamic conditions. Comparison of sludge properties and cake layer structure from the two systems was made to elucidate major factors governing the different filtration characteristics. There were more soluble microbial products (SMP) and biopolymer clusters (BPC) produced and a larger portion of fine flocs (<15 μm) in the thermophilic SAnMBR. Analysis of bound extracellular polymeric substances (EPS) showed that the thermophilic sludge had a higher protein/polysaccharide ratio in EPS, as compared to that in the mesophilic sludge. A series of analyses, including Fourier transform infrared (FTIR) spectroscopy, energy dispersive X-ray spectroscopy (EDX), confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), atomic force microscopy (AFM) and particle size analyzer showed that the cake layer formed in the thermophilic SAnMBR contained higher levels of both organic and inorganic foulants, smaller particle sizes, and especially, a denser and more compact sludge cake structure. These results indicate that floc size, SMP, BPC, bound EPS as well as cake layer structure are the major factors governing membrane fouling in SAnMBR systems.

Dry anaerobic ammonia–methane production from chicken manure

The effect of temperature on production of ammonia during dry anaerobic fermentation of chicken manure (CM), inoculated with thermophilic methanogenic sludge, was investigated in a batch condition for 8 days. Incubation temperature did not have a significant effect on the production of ammonia. Almost complete inhibition of production of methane occurred at 55 and 65 degrees C while quite low yields of 8.45 and 6.34 ml g(-1) VS (volatile solids) were observed at 35 and 45 degrees C due to a higher accumulation of ammonia. In order to improve the production of methane during dry anaerobic digestion of CM, stripping of ammonia was performed firstly on the CM previously fermented at 65 degrees C for 8 days: the stripping for 1 day at 85 degrees C and pH 10 removed 85.5% of ammonia. The first-batch fermentation of methane for 75 days was conducted next, using the ammonia-stripped CM inoculated with methanogenic sludge at different ratios, (CM: thermophilic sludge) of 1:2, 1:1, and 2:1 on volume per volume basis at both 35 and 55 degrees C. Production of methane improved and was higher than that of the control (without stripping of ammonia) but the yield of 20.4 ml g(-1) VS was still low, so second stripping of ammonia was conducted, which resulted in 74.7% removal of ammonia. A great improvement in the production of methane of 103.5 ml g(-1) VS was achieved during the second batch for 55 days.

Characteristics of thermophilic granulation processes in UASB reactors

Thermophilic granulation processes were examined with synthetic wastewaters and were compared with the parallel mesophilic ones. The results indicated that granules could be formed well under compatible thermophilic surroundings in both carbohydrate- and protein-rich substrates. Compared with the mesophilic process, thermophilic granulation took relatively longer time, but thermophilic UASB reactors had much higher treatment capacity and efficiency than the mesophilic ones. Under the same running conditions, thermophilic sludge had lower contents of Extracellular Polymer (ECP) than mesophilic ones, which might be one of the reasons of the longer period needed for thermophilic granulation.

A two‐stage process for simultaneous thermophilic sludge digestion, pathogen control and metal leaching

A consortium of aerobic sulphur oxidizing microorganisms native to municipal sewage sludge has been developed that is capable of reducing the pH of sludge from 7.0 to 2.0 under mild‐thermophilic (45 °C) temperatures. This mild‐thermophilic, acidophilic acclimatized consortium has then been used to develop a thermophilic acidophilic culture capable of reducing the pH of municipal sewage sludge from 7.0 to 4.8 at 60 °C. Furthermore, these two cultures have been used to develop a novel two‐stage aerobic process functioning under thermophilic and mild‐thermophilic conditions capable of simultaneous sludge digestion, pathogen control and metal leaching. Studies with 1.5 L reactors indicate that at the end of the process, concentration of volatile suspended solids has stabilized, no coliforms are present, and 90–98% copper, 90–94% manganese and 95–100% zinc has been leached.

Sulfate reduction at pH 4 during the thermophilic (55°C) acidification of sucrose in UASB reactors

Continuous sulfate reduction at pH 4.0 was demonstrated in a pH controlled thermophilic (55 degrees C) upflow anaerobic sludge bed reactor fed with sucrose at a COD/SO(4)(2-) ratio of 0.9 and an organic loading rate of 0.8 and 1.9 gCOD (l(reactor) d)(-1) for a period of 78 days. A nearly complete sulfate reduction efficiency was achieved throughout the reactor run, corresponding to sulfate removal rates of 0.91 and 1.92 g (l(reactor) d)(-1) at sulfate loading rates of 0.94 and 2 g (l(reactor) d)(-1), respectively, by keeping the sulfide concentration below 20 mg l(-1) due to stripping with nitrogen gas. Acidification was always complete and acetate was the only degradation intermediate left in the effluent, which did not exceed 180 mgCOD l(-1) in pseudo-stationary states. The sludge was well retained in the reactor and kept its granular form. Zn, Cu, Se, and Mo accumulated in the sludge, whereas Co, Ni, Fe, and Mn leached from the sludge, despite their continuous supply to the reactor via the influent. The bacterial diversity in the reactor sludge at the end of the reactor run was low and the culture was dominated by one acidifying species, resembling Thermoanaerobacterium sp., and one sulfate reducing species, resembling Desulfotomaculum sp.

Thermodesulfovibrio aggregans sp. nov. and Thermodesulfovibrio thiophilus sp. nov., anaerobic, thermophilic, sulfate-reducing bacteria isolated from thermophilic methanogenic sludge, and emended description of the genus Thermodesulfovibrio

Four obligately anaerobic, thermophilic, sulfate-reducing bacterial strains, designated TGE-P1(T), TDV(T), TGL-LS1 and TSL-P1, were isolated from thermophilic (operated at 55 degrees C) methanogenic sludges from waste and wastewater treatment. The optimum temperature for growth of all the strains was in the range 55-60 degrees C. The four strains grew by reduction of sulfate with a limited range of electron donors, such as hydrogen, formate, pyruvate and lactate. In co-culture with the hydrogenotrophic methanogen Methanothermobacter thermautotrophicus DeltaH(T), strains TGE-P1(T), TGL-LS1 and TSL-P1 were able to utilize lactate syntrophically for growth. The DNA G+C contents of all the strains were in the range 34-35 mol%. The major cellular fatty acids of the strains were iso-C(17 : 0), iso-C(16 : 0), C(16 : 0) and anteiso-C(15 : 0). Phylogenetic analyses based on 16S rRNA gene sequences revealed that the strains belong to the Thermodesulfovibrio clade of the phylum 'Nitrospirae'. On the basis of their physiological, chemotaxonomic and genetic properties, strains TGL-LS1 (=JCM 13214) and TSL-P1 (=JCM 13215) were classified as strains of Thermodesulfovibrio islandicus. Two novel species of the genus Thermodesulfovibrio are proposed to accommodate the other two isolates: Thermodesulfovibrio aggregans sp. nov. (type strain TGE-P1(T) =JCM 13213(T) =DSM 17283(T)) and Thermodesulfovibrio thiophilus sp. nov. (type strain TDV(T) =JCM 13216(T) =DSM 17215(T)). To examine the ecological aspects of Thermodesulfovibrio-type cells in the sludge from which the strains were originally isolated, an oligonucleotide probe targeting 16S rRNA of all Thermodesulfovibrio species was designed and applied to thin sections of thermophilic sludge granules. Fluorescence in situ hybridization using the probe revealed rod- or vibrio-shaped cells as a significant population within the sludge, indicating their important role in the original ecosystem.

Evaluating limiting steps of anaerobic degradation of food waste based on methane production tests

This research adapted a batch test for biochemical methane production (BMP) to follow the degradation of complex compounds such as proteins and vegetable oils. The test measured the transformation of albumin and olive oil into methane under mesophilic and thermophilic conditions and assess limiting step in the overall degradation process. The thermophilic sludge used for the BMP tests was adapted during ten month from mesophilic sludge while being fed food waste. As compared to acetic acid, the specific rate of transformation of albumin and olive oil into methane reached 22 and 51%, respectively, under mesophilic conditions. Acetoclastic methanogenesis was not the limiting step in the presence of albumin or olive oil (and its monomer-like molecules such as amino acids, glycerol and oleic acid). Rather, the degradation of albumin was restricted by the presence of proteins. The thermophilically adapted sludge showed good proteolytic activity, but its acetoclastic methanogens were unable to degrade olive oil, because of the inhibitory effect of oleic acid.

Development and optimisation of VFA driven DEAMOX process for treatment of strong nitrogenous anaerobic effluents

The recently proposed DEAMOX (DEnitrifying AMmonium OXidation) process combines the anammox reaction with autotrophic denitrifying conditions using sulphide as an electron donor for the production of nitrite from nitrate within an anaerobic biofilm. This paper firstly presents a feasibility study of the DEAMOX process using synthetic (ammonia + nitrate) wastewater where sulphide is replaced by volatile fatty acids (VFA) as a more widespread electron donor for partial denitrification. Under the influent N-NH+4/N-NO3(-) and COD/N-NO3(-) ratios of 1 and 2.3, respectively, the typical efficiencies of ammonia removal were around 40% (no matter whether a VFA mixture or only acetate were used) for nitrogen loading rates (NLR) up to 1236 mg N/l/d. This parameter increased to 80% by increasing the influent COD/N-NO3(-) ratio to 3.48 and decreasing the influent N-NH4 +/N-NO3(-) ratio to 0.29. As a result, the total nitrogen removal increased to 95%. The proposed process was further tested with typical strong nitrogenous effluent such as reject water (total N, 530-566 mg N/l; total COD, 1530-1780 mg/l) after thermophilic sludge anaerobic digestion. For this, the raw wastewater was split and partially ( approximately 50%) fed to a nitrifying reactor (to generate nitrate) and the remaining part ( approximately 50%) was directed to the DEAMOX reactor where this stream was mixed with the nitrified effluent. Stable process performance up to NLR of 1,243 mg N/l/d in the DEAMOX reactor was achieved resulting in 40, 100, and 66% removal of ammonia, NOx(-), and total nitrogen, respectively.

Diversity of anaerobic microorganisms involved in long-chain fatty acid degradation in methanogenic sludges as revealed by RNA-based stable isotope probing.

Long-chain fatty acid (LCFA) degradation is a key step in methanogenic treatment of wastes/wastewaters containing high concentrations of lipids. However, despite the importance of LCFA-degrading bacteria, their natural diversity is little explored due to the limited availability of isolate information and the lack of appropriate molecular markers. We therefore investigated these microbes by using RNA-based stable isotope probing. We incubated four methanogenic sludges (mesophilic sludges MP and MBF and thermophilic sludges TP and JET) with (13)C-labeled palmitate (1 mM) as a substrate. After 8 to 19 days of incubation, we could detect (13)C-labeled bacterial rRNA. A density-resolved terminal restriction fragment length polymorphism fingerprinting analysis showed distinct bacterial populations in (13)C-labeled and unlabeled rRNA fractions. The bacterial populations in the (13)C-labeled rRNA fractions were identified by cloning and sequencing of reverse-transcribed 16S rRNA. Diverse phylogenetic bacterial sequences were retrieved, including those of members of the family Syntrophaceae, clone cluster MST belonging to the class Deltaproteobacteria, Clostridium clusters III and IV, phylum Bacteroidetes, phylum Spirochaetes, and family Syntrophomonadaceae. Although Syntrophomonadaceae species are considered to be the major fatty acid-degrading syntrophic microorganisms under methanogenic conditions, they were detected in only two of the clone libraries. These results suggest that phylogenetically diverse bacterial groups were active in situ in the degradation of LCFA under methanogenic conditions.

Characterisation of community structure of bacteria in parallel mesophilic and thermophilic pilot scale anaerobe sludge digesters

The aim of the present work was to compare the microbial communities of a mesophilic and a thermophilic pilot scale anaerobe sludge digester. For studying the communities cultivation independent chemotaxonomical methods (RQ and PLFA analyses) and T-RFLP were applied. Microbial communities of the mesophilic and thermophilic pilot digesters showed considerable differences, both concerning the species present, and their abundance. A Methanosarcina sp. dominated the thermophilic, while a Methanosaeta sp. the mesophilic digester among Archaea. Species diversity of Bacteria was reduced in the thermophilic digester. Based on the quinone patterns in both digesters the dominance of sulphate reducing respiratory bacteria could be detected. The PLFA profiles of the digester communities were similar though in minor components characteristic differences were shown. Level of branched chain fatty acids is slightly lower in the thermophilic digester that reports less Gram positive bacteria. The relative ratio of fatty acids characteristic to Enterobacteriaceae, Bacteroidetes and Clostridia shows differences between the two digesters: their importance generally decreased under thermophilic conditions. The sulphate reducer marker (15:1 and 17:1) fatty acids are present in low quantity in both digesters.

Connection between oxygen uptake rate and carbon dioxide evolution rate in aerobic thermophilic sludge digestion

The main aim of studying the relation of carbon dioxide evolution rate CER to oxygen uptake rate (OUR) is the possible application of CER in mathematical modelling of aerobic biodegradation processes instead of OUR. Biodegradation tests using glucose and sewage sludge as feed were performed to compare the OUR and CER. The respiratory quotient (RQ) was 0.9 mol CO2 &times; mol O2-1 in endogenous stage while its value was increased to 1.2 mol CO2 &times; mol O2-1 during glucose degradation. At higher F/M ratios and high respiration rates RQ values up to 2.95 were observed which may indicate the appearance of anaerobic degradation pathways. These results prove that there is no direct, simple relation between OUR and CER in case of sewage sludge degradation so direct substitution of OUR with CER in modelling studies is not feasible.

Population dynamics of anaerobic microbial consortia in thermophilic methanogenic sludge treating paper-containing solid waste

To maintain a stable thermophilic (55 degrees C) anaerobic digestion treating toilet paper-containing garbage, it is necessary to operate the digester at long hydraulic retention time (HRT) and low organic loading rate (OLR). Critical conditions of the digestion were investigated by operating the digester at HRT 23 days and OLR 3.4 gCOD(Cr)/L/d (R1) or HRT 14 days and OLR 5.6 gCOD/Cr)/L/d (R2) separately. Characteristics of methanogenesis of the two digesters were examined by measuring gas generation volume and volatile fatty acids (VFA) concentration, and the populations of four anaerobic acidogens and three methanogens were analyzed using quantitative PCR method. In digester R1, methanogenic activity was unstable but it could be recovered by stopping feeding as though VFA accumulation occurred. The population of acidogens and two methanogens were maintained at 10(11) - 10(13) copies/L, however, the population of Methanoculleus could not be recovered after methanogenesis recovering. In digester R2, the period of methanogenesis was significantly shorter than that in digester R1. Both the acidogens and the methanogens could not be maintained at a stable concentration. It is suggested that the critical HRT to sustain the population of acidogens in this process should be longer than 14 days and for all kinds of methanogens, HRT should be longer than 23 days.

Performance and methanogenic community of rotating disk reactor packed with polyurethane during thermophilic anaerobic digestion

A newly developed anaerobic rotating disk reactor (ARDR) packed with polyurethane was used in continuous mode for organic waste removal under thermophilic (55 °C) anaerobic conditions. This paper reports the effects of the rotational speed on the methanogenic performance and community in an ARDR supplied with acetic acid synthetic wastewater as the organic substrate. The best performance was obtained from the ARDR with the rotational speed ( ω) of 30 rpm. The average removal of dissolved organic carbon was 98.5%, and the methane production rate was 393 ml/l-reactor/day at an organic loading rate of 2.69 g/l-reactor/day. Under these operational conditions, the reactor had a greater biomass retention capacity and better reactor performance than those at other rotational speeds (0, 5 and 60 rpm). The results of 16S rRNA phylogenetic analysis indicated that the major methanogens in the reactor belonged to the genus Methanosarcina spp. The results of real-time polymerase chain reaction (PCR) analysis suggested that the cell density of methanogenic archaea immobilized on the polyurethane foam disk could be concentrated more than 2000 times relative to those in the original thermophilic sludge. Scanning electron microphotographs showed that there were more immobilized microbes at ω of 30 rpm than 60 rpm. A rotational speed on the outer layer of the disk of 6.6 m/min could be appropriate for anaerobic digestion using the polyurethane ARDR.

Microbial activity and population structure of anaerobic sludge alternately exposed to mesophilic and thermophilic conditions

The microbial community structures of anaerobic sludge, alternately exposed to mesophilic and thermophilic condition, were investigated and their microbial activities under mesophilic and thermophilic conditions. PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) profiles for the eubacterial and archaebacterial communities from thermophilic sludge alternately exposed to mesophilic condition (TSEM) and mesophilic sludge alternately exposed to thermophilic condition (MSET) had a few populations in common, these are probably thermophilic microorganisms. The population profiles for archaebacterial communities of TSEM and MSET were quite different from each other. For TSEM under thermophilic condition, the specific methanogenic activity (SMA) was 159.6mL CH4/g VSS/d, which was higher than 106.3mL CH4/g VSS/d under mesophilic condition. The SMA for MSET under both temperature conditions was as much as those of TSEM. The lag phase period for MSET in the batch culture was around 2 times longer than TSEM, but the lag periods for both sludges under thermophilic condition were shorter than those under mesophilic condition. The acidogenic activity for both sludge types in MSET was slightly higher than TSEM under the thermophilic and mesophilic conditions. The specific hydrolytic activity for the two sludge types was higher under the mesophilic condition, and the hydrolysis potential of TSEM incubated under mesophilic condition was higher than other cases.

Identification of mesophilic and thermophilic fermentative species in anaerobic granular sludge

Large quantities of biodegradable food waste in the form of fruit and vegetables are still being deposited in landfill sites in Ireland. The development of an anaerobic digestion process using fermentative species which degrade the carbohydrate-rich waste could divert the food waste from landfills. We identified fermentative species grown on glucose and sucrose at mesophilic and thermophilic temperatures using molecular biology techniques. The dominating fermentative bacteria of the mesophilic sludge were of the Bacteroidetes and Spirochaetes classes. Although both groups of bacteria are typically fermentative their substrate range appears to be limited. The dominating fermentative bacteria in the thermophilic sludge was Thermoanaerobacterium aotearoense of the Clostridia class. The indications are that Thermoanaerobacterium aotearoense may be highly suitable to biodegrade a carbohydrate-rich influent feed due to its possibly very rapid growth rate and also an extensive substrate range.

Anaerobic treatment of phenol in wastewater under thermophilic condition

Over 99% of phenol was effectively degraded in an upflow anaerobic sludge blanket (UASB) reactor at 55 °C with 40 h of hydraulic retention time (HRT) for a wastewater containing 630 mg/L of phenol, corresponding to 1500 mg/L of chemical oxygen demand (COD) and a loading rate of 0.9 g-COD/L/d. The maximum specific methanogenic activity (SMA) of the phenol-degrading sludge was 0.09 g-CH 4-COD/g-volatile suspended solids (VSS)/d. Based on 16S rDNA analysis, a total of 21 operational taxonomy units (OTUs) were found in the sludge, of which eight (42.6% of the total population) were related to the sequences in the GenBank with similarity of over 97%, and 13 (79.6%) were affiliated with the known thermophilic species. Additional SMA data and phylogenetic analysis suggest that the degradation pathway of phenol for thermophilic sludge was likely via caproate, instead of benzoate as for the mesophilic sludge.

混合微生物群によるでんぷんの高温水素発酵特性

The effects of inoculum on the characteristics of thermophilic hydrogen fermentation were investigated by performing a long-term continuous experiment lasting more 50 days. Continuous hydrogen fermentation of starch was conducted under thermophilic condition (55°C) by using five types of inoculum, including thermophilically digested sludge of activated sludge, cattle manure and sewage sludge, compost, and thermophilic acidogenic sludge. Five chemostat-type reactors were operated at an HRT (hydraulic retention time) of 24h. Stable hydrogen production was achieved in each culture without any methane production during the long-term continuous experiment. The highest hydrogen yield (2.32 mol H2/mol hexose converted) was obtained by thermophilically digested sludge of activated sludge. The thermophilic hydrogen fermentation of starch by thermophilically digested sludge of activated sludge was expressed using the following stoichiometric equation : starch → 2.32n H2 + 2.14n CO2 + 0.5n acetate + 0.63n butyrate + 0.11n biomass (C5H9O3N).

Diversity, Localization, and Physiological Properties of Filamentous Microbes Belonging to Chloroflexi Subphylum I in Mesophilic and Thermophilic Methanogenic Sludge Granules

We previously reported that the thermophilic filamentous anaerobe Anaerolinea thermophila, which is the first cultured representative of subphylum I of the bacterial phylum Chloroflexi, not only was one of the predominant constituents of thermophilic sludge granules but also was a causative agent of filamentous sludge bulking in a thermophilic (55 degrees C) upflow anaerobic sludge blanket (UASB) reactor in which high-strength organic wastewater was treated (Y. Sekiguchi, H. Takahashi, Y. Kamagata, A. Ohashi, and H. Harada, Appl. Environ. Microbiol. 67:5740-5749, 2001). To further elucidate the ecology and function of Anaerolinea-type filamentous microbes in UASB sludge granules, we surveyed the diversity, distribution, and physiological properties of Chloroflexi subphylum I microbes residing in UASB granules. Five different types of mesophilic and thermophilic UASB sludge were used to analyze the Chloroflexi subphylum I populations. 16S rRNA gene cloning-based analyses using a 16S rRNA gene-targeted Chloroflexi-specific PCR primer set revealed that all clonal sequences were affiliated with the Chloroflexi subphylum I group and that a number of different phylotypes were present in each clone library, suggesting the ubiquity and vast genetic diversity of these populations in UASB sludge granules. Subsequent fluorescence in situ hybridization (FISH) of the three different types of mesophilic sludge granules using a Chloroflexi-specific probe suggested that all probe-reactive cells had a filamentous morphology and were widely distributed within the sludge granules. The FISH observations also indicated that the Chloroflexi subphylum I bacteria were not always the predominant populations within mesophilic sludge granules, in contrast to thermophilic sludge granules. We isolated two mesophilic strains and one thermophilic strain belonging to the Chloroflexi subphylum I group. The physiological properties of these isolates suggested that these populations may contribute to the degradation of carbohydrates and other cellular components, such as amino acids, in the bioreactors.

Effect of solids retention time (SRT) on pilot scale anaerobic thermophilic sludge digestion

This paper describes anaerobic thermophilic sludge digestion (55 °C) in a continuously stirred tank reactor (CSTR) on a pilot-plant scale (150 L). The experimental protocol was defined to examine the effect of the increase in the organic loading rate on the efficiency of the digester and to report on its steady-state performance. The reactor was subjected to a programme of steady-state operation over a range of solids retention times (SRTs) of 75, 40, 27, 20 and 15 days and organic loading rates (OLR) in the range 0.4–2.2 kg VS/(m 3 day). The digester was fed with raw sludge (containing approximately 35 kg/m 3 volatile solids (VS)) once daily during the 75-day SRT period, twice daily during the 40-day SRT period and three times a day during the 27-, 20- and 15-day SRT periods. The reactor was initially operated with an organic loading rate of 0.4 kg VS/(m 3 day) and an SRT of 75 days. The volatile solids removal efficiency in the reactor was found to be 73%, while the volumetric methane production rate produced in the digester reached 0.02 m 3/(m 3 day). Over a 338-day operating period, an OLR of 2.2 kg VS/(m 3 day) was achieved with 49.1% VS removal efficiency in the pilot sludge digester, at which time the volumetric methane production rate content of biogas produced in the digester reached 0.4 m 3/(m 3 day). The chemical oxygen demand (COD) mass balance obtained indicated that COD used for methane generation increased when the SRT was decreased or when the influent organic loading rate was increased. This implies that the amount of COD used in the anabolism route decreased with SRT due the microbial population becoming adapted to new operational conditions and more COD being used to generate methane.