The effect of temperature variation on biomethanation at high altitude

The aim of the current study was to examine effects of daily temperature variations on the performance of anaerobic digestion. Forced square-wave temperature variations (between 11 and 25, 15 and 28, and 19 and 32 degrees C) were imposed on a bench-scale digester using a mixture of llama-cow-sheep manure in a semi-continuous process. The volumetric biogas production rate, methane yield, and the volatile solid reductions were compared with the results obtained from anaerobic digestion (AD) at constant temperatures. The forced cyclic variations of temperature caused large cyclic variations in the rate of gas production and the methane content. As much as 94-97% of the daily biogas was obtained in the 12h half-cycle at high temperature. The values for volumetric biogas production rate and methane yield increased at higher temperatures. The average volumetric biogas production rate for cyclic operation between 11 and 25 degrees C was 0.22Ld(-1)L(-1) with a yield of 0.07m3CH4kg(-1) VS added (VSadd), whereas for operation between 15 and 29 degrees C the volumetric biogas production rate increased by 25% (to 0.27Ld(-1)L(-1) with a yield of 0.08m3CH4kg(-1) VSadd). In the highest temperature region a further increase of 7% in biogas production was found and the methane yield was 0.089m(3)CH(4)kg(-1) VSadd. The employed digester showed an immediate response when the temperature was elevated, which indicates a well-maintained metabolic capacity of the methanogenic bacteria during the period of low temperature. Overall, periodic temperature variations appear to give less decrease in process performance than a priori anticipated.     

Quantifying electricity generation and waste transformations in a low-cost, plug-flow anaerobic digestion system

Methane production, electricity production, and wastewater transformations were quantified for a digestion system that combines biogas from a swine digester and dairy digester in Costa Rica. The low-cost, plug-flow digesters were not heated and were operated in the lower portion of the mesophilic range (25–27 °C).The dairy digester produced 27.5 m³/day of biogas with 62.6% methane and reduced organic matter (COD) by 86%. The swine digester produced 6.0 m³/day of biogas with 76.4% methane and reduced COD by 92%. Combining biogas from a swine and dairy digester, increased electricity production due to the higher biogas production rate of the dairy farm and the higher quality biogas obtained from the swine farm. The farm’s 2-h peak electricity demand (12.9 kW/day) was 81.8% met. The electricity was produced using manure equivalent to the quantity excreted by 5 dairy cows and 40 pigs remaining in corrals 100% of the time.The $21,000 capital cost of the digester project will be recovered in 10.1 years through electricity savings and reductions in wastewater fines. If the generator were more appropriately sized for the farm, the capital recovery time would have been 7.6 years.     

Effects of solid-liquid separation on recovering residual methane and nitrogen from digested dairy cow manure

The feasibility of optimizing methane and nitrogen recovery of samples obtained from farm biogas digester (35 degrees C) and post-storage tank (where digested material is stored for 9-12 months) was studied by separating the materials into different fractions using 2, 1, 0.5 and 0.25 mm sieves. Mass-balances revealed that digested material mainly consists of <0.25 mm (60-69%) and >2 mm (18-27%) fractions, while fractions between 2 and 0.2 mm made the rest. Incubation of solid fractions >0.25 mm of digester material at 35 degrees C resulted in specific methane yields of 0.060-0.085 m(3)kg(-1) volatile solids (VS) during initial 30-50 d and 0.16-0.18 m(3)kg(-1)VS at the end of 340 d incubation. Similarly, fractions >0.25 mm of post-storage tank material produced 0.055-0.092 m(3)kg(-1)VS and 0.13-0.16 m(3)kg(-1)VS of methane after 30-50 d and after 250 d, respectively. Methane yields for fractions <0.25 mm of post-storage tank was 0.03 m(3)kg(-1)VS after 30-50 d and 0.05 m(3)kg(-1)VS after 250 d compared to 0.20 m(3)kg(-1)VS and 0.41 m(3)kg(-1)VS, respectively for the same fraction of digester material. Separation of digested cow manure into solids and liquid fractions to recover methane may be feasible only for post-storage tank material and not for digester material. Nitrogen management would not be feasible with neither material as total nitrogen and ammonium-nitrogen concentrations were equally distributed among the segregated fractions.     

Effect of anaerobic digestion temperature on odour, coliforms and chlortetracycline in swine manure or monensin in cattle manure

Aims: This study evaluated the effect of anaerobic digestion at 22, 38 and 55°C on odour, coliforms and chlortetracycline (CTC) in swine manure or monensin (MON) in cattle manure. Methods and Results: Swine or cattle were fed the respective growth promotant, manure was collected, and 2‐l laboratory methane digesters were established at the various temperatures and sampled over 25 or 28 days. After 21 days, the concentration of CTC in the 22, 38 and 55°C swine digester slurries decreased 7, 80 and 98%, respectively. Coliforms in the 22°C digester slurries were still viable after 25 days; however, they were not detectable in the 38 and 55°C slurries after 3 and 1 days, respectively. After 28 days, the concentration of MON in the 22, 38 and 55°C cattle digester slurries decreased 3, 8 and 27%, respectively. Coliforms in the 22°C cattle digester slurries were still viable after 28 days; however, they were not detectable in the 38 and 55°C slurries after 14 and 1 days, respectively. Conclusions: These studies indicate that anaerobic digestion at 38 or 55°C may be an effective treatment to reduce coliforms and CTC; however, it is not an effective treatment to reduce MON. Significance and Impact of the Study: More studies are needed to determine which pharmaceuticals are susceptible to degradation by a specific manure treatment to prevent negative environmental consequences.     

Anaerobic digestion of animal waste: waste strength versus impact of mixing

We studied the effect of mode of mixing (biogas recirculation, impeller mixing, and slurry recirculation) and waste strength on the performance of laboratory scale digesters. The digesters were fed with 5% and 10% manure slurry, at a constant energy supply per unit volume (8 W/m3). The experiments were conducted in eight laboratory scale digesters, each having a working volume of 3.73 L, at a controlled temperature of 35+/-2 degrees C. Hydraulic retention time (HRT) was kept constant at 16.2 days, resulting in a total solids (TS) loading rate of 3.08 g/Ld and 6.2 g/Ld for 5% and 10% manure slurry feeds, respectively. Results showed that the unmixed and mixed digesters performed quite similarly when fed with 5% manure slurry and produced biogas at a rate of 0.84-0.94 L/Ld with a methane yield of 0.26-0.31 L CH4/g volatile solids (VS) loaded. This was possibly because of the low solids concentration in the case of 5% manure slurry, where mixing created by the naturally produced gas might be sufficient to provide adequate mixing. However, the effect of mixing and the mode of mixing became prominent in the case of the digesters fed with thicker manure slurry (10%). Digesters fed with 10% manure slurry and mixed by slurry recirculation, impeller, and biogas recirculation produced approximately 29%, 22% and 15% more biogas than unmixed digester, respectively. Deposition of solids inside the digesters was not observed in the case of 5% manure slurry, but it became significant in the case of 10% manure slurry. Therefore, mixing issue becomes more critical with thicker manure slurry.     

Effect of feed to inoculum ratios on biogas yields of food and green wastes

Biogas and methane yields of food and green wastes and their mixture were determined using batch anaerobic digesters at mesophilic (35 ± 2 °C) and thermophilic (50 ± 2 °C) temperatures. The mixture was composed of 50% food waste and 50% green waste, based on the volatile solids (VS) initially added to the reactors. The thermophilic digestion tests were performed with four different feed to inoculum (F/I) ratios (i.e., 1.6, 3.1, 4.0 and 5.0) and the mesophilic digestion was conducted at one F/I (3.1). The results showed that the F/I significantly affected the biogas production rate. At four F/Is tested, after 25 days of thermophilic digestion, the biogas yield was determined to be 778, 742, 784 and 396 mL/g VS for food waste, respectively; 631, 529, 524 and 407 mL/g VS for green waste, respectively; and 716, 613, 671 and 555 mL/g VS for the mixture, respectively. About 80% of the biogas production was obtained during the first 10 days of digestion. At the F/I of 3.1, the biogas and methane yields from mesophilic digestion of food waste, green waste and their mixture were lower than the yields obtained at thermophilic temperature. The biogas yields were 430, 372 and 358 mL/g VS, respectively, and the methane yields were 245, 206, and 185 mL/g VS, respectively.     

Methane production using whole and screened dairy manure in conventional and fixed-film reactors

The technical feasibility of adopting the fixed-film reactor concept for biogas production from screened dairy manure was investigated. The methane production capability of laboratory-scale 4-L anaerobic reactors (conventional and fixed-film) receiving screened dairy manure and operated at 35 degrees C was compared. Dairy manure filtrate with 4.4% total solids (TS) and 3.4% volatile solids (VS) (average value) was prepared from 1:1 manure-water slurry. The feed material was added intermittently at loading rates ranging from 2.34 to 25 and 2.25 to 785 g VS/L d, respectively, for the conventional and fixed-film reactors. Maximum methane production rate (L CH(4)/L d) for the conventional reactor was 0.63 L CH(4)/L d achieved at a 6-day hydraulic retention time (HRT). For the fixed-film reactor the maximum production rate was 3.53 L CH(4)/L d when operated at a loading rate of 262 g VS/L d (3 h HRT). The fixed-film reactor was capable of sustaining a loading of 785 g VS/L d (1 h HRT). The fixed-film reactor performed much better than the conventional reactors. These results indicate that a large reduction of required reactor volume is possible through application of a fixed-film concept combined with a liquid-solid separation pretreatment of dairy manure.     

Comparative evaluation of biogas production from dairy manure and co‐digestion with maize silage by CSTR and new anaerobic hybrid reactor

This study aimed to investigate potential methane production through anaerobic digestion of dairy manure and co‐digestion with maize silage. Two different anaerobic reactor configurations (single‐stage continuously stirred tank reactor [CSTR] and hybrid anaerobic digester) were used and biogas production performances for each reactor were compared. The HR was planned to enable phase separation in order to improve process stability and biogas production under higher total solids loadings (≥4%). The systems were tested under six different organic loading rates increased steadily from 1.1 to 5.4 g VS/L.d. The CSTR exhibited lower system stability and biomass conversion efficiency than the HR. The specific biogas production of the hybrid system was between 440 and 320 mL/gVS with 81–65% volatile solids (VS) destruction. The hybrid system provided 116% increase in specific biogas production and VS destruction improved by more than 14%. When MS was co‐digested together with dairy manure, specific biogas production rates increased about 1.2‐fold. Co‐digestion was more beneficial than mono‐material digestion. The hybrid system allowed for generating methane enriched biogas (>75% methane) by enabling phase separation in the reactor. It was observed that acidogenic conditions prevailed in the first two compartments and the following two segments as methanogenic conditions were observed. The pH of the acidogenic part ranged between 4.7 and 5.5 and the methanogenic part was between 6.8 and 7.2.     

Influence of temperature on Biogas Production from Pistia stratiotes

Pistia stratiotes, an aquatic weed, was investigated as a substrate for biogas production. Experiments were carried out as batch runs in laboratory-scale digesters with the addition of inoculum (digested cattle manure). Gas yields were in the range of 533–707 litres kg⁻¹ VS (STP), respectively, 21–28 litres kg⁻¹ fresh weight of P. stratiotes, with 30 days digestion time at temperatures of 29·5, 33·0 and 37·5°C. The average methane content was 58–68%. Due to its high biodegradability (approximately 83–99% of VS) Pistia stratiotes is very suitable as a substrate for biogas production.     

Optimisation of biogas production from manure through serial digestion: lab-scale and pilot-scale studies

In the present study, the possibility of optimizing biogas production from manure by serial digestion was investigated. In the lab-scale experiments, process performance and biogas production of serial digestion, two methanogenic continuously stirred tank reactors (CSTR) connected in series, was compared to a conventional one-step CSTR process. The one-step process was operated at 55 degrees C with 15d HRT and 5l working volume (control). For serial digestion, the total working volume of 5l was distributed as 70/30%, 50/50%, 30/70% or 13/87% between the two methanogenic reactors, respectively. Results showed that serial digestion improved biogas production from manure compared to one-step process. Among the tested reactor configurations, best results were obtained when serial reactors were operated with 70/30% and 50/50% volume distribution. Serial digestion at 70/30% and 50/50% volume distribution produced 13-17.8% more biogas and methane and, contained low VFA and residual methane potential loss in the effluent compared to the one-step CSTR process. At 30/70% volume distribution, an increase in biogas production was also noticed but the process was very unstable with low methane production. At 13/87% volume distribution, no difference in biogas production was noticed and methane production was much lower than the one-step CSTR process. Pilot-scale experiments also showed that serial digestion with 77/23% volume distribution could improve biogas yields by 1.9-6.1% compared to one-step process. The study thus suggests that the biogas production from manure can be optimized through serial digestion with an optimal volume distribution of 70/30% or 50/50% as the operational fluctuations are typically high during full scale application. However, process temperature between the two methanogenic reactors should be as close as possible in order to derive the benefits of serial coupling.     

Novel membrane bioreactor: Able to cope with fluctuating loads, poorly water soluble VOCs, and biomass accumulation

We determined the effect of different mixing intensities on the performance, methanogenic population dynamics, and juxtaposition of syntrophic microbes in anaerobic digesters treating cow manure from a dairy farm. Computer automated radioactive particle tracking in conjunction with computational fluid dynamics was performed to quantify the shear levels locally. Four continuously stirred anaerobic digesters were operated at different mixing intensities of 1,500, 500, 250, and 50 revolutions per min (RPM) over a 260-day period at a temperature of 34 +/- 1 degrees C. Animal manure at a volatile solids (VS) concentration of 50 g/L was fed into the digesters daily at five different organic loading rates between 0.6 and 3.5 g VS/L day. The different mixing intensities had no effect on the biogas production rates and yields at steady-state conditions. A methane yield of 0.241 +/- 0.007 L CH(4)/g VS fed was obtained by pooling the data of all four digesters during steady-state periods. However, digester performance was affected negatively by mixing intensity during startup of the digesters, with lower biogas production rates and higher volatile fatty acids concentrations observed for the 1,500-RPM digester. Despite similar methane production yields and rates, the acetoclastic methanogenic populations were different for the high- and low-intensity mixed digesters with Methanosarcina spp. and Methanosaeta concilii as the predominant methanogens, respectively. For all four digesters, epifluorescence microscopy revealed decreasing microbial floc sizes beginning at week 4 and continuing through week 26 after which no microbial flocs remained. This decrease in size, and subsequent loss of microbial flocs did not, however, produce any long-term upsets in digester performance.     

Characterization of food waste as feedstock for anaerobic digestion

Food waste collected in the City of San Francisco, California, was characterized for its potential for use as a feedstock for anaerobic digestion processes. The daily and weekly variations of food waste composition over a two-month period were measured. The anaerobic digestibility and biogas and methane yields of the food waste were evaluated using batch anaerobic digestion tests performed at 50 degrees C. The daily average moisture content (MC) and the ratio of volatile solids to total solids (VS/TS) determined from a week-long sampling were 70% and 83%, respectively, while the weekly average MC and VS/TS were 74% and 87%, respectively. The nutrient content analysis showed that the food waste contained well balanced nutrients for anaerobic microorganisms. The methane yield was determined to be 348 and 435 mL/gVS, respectively, after 10 and 28 days of digestion. The average methane content of biogas was 73%. The average VS destruction was 81% at the end of the 28-day digestion test. The results of this study indicate that the food waste is a highly desirable substrate for anaerobic digesters with regards to its high biodegradability and methane yield.     

Psychrophilic anaerobic digestion biotechnology for swine mortality disposal

The objective of this study was to investigate the feasibility of using psychrophilic anaerobic digestion in sequencing batch reactors (PADSBRs) to co-digest grinded swine carcasses and swine manure slurry at 20 degrees C and 25 degrees C. The PADSBRs were operated on two-week and four-week treatment cycle lengths, which included the fill, react, and draw phases. Two carcass loading rates (CLRs) were tested, that is 20 and 40g of carcass per litre of manure, which were equivalent to 4 and 8 times, respectively, the normal mortality rate on commercial farms. The PADSBR performance was compared to that of PADSBRs operated at 25 degrees C and fed manure only. The addition of swine carcass to PADSBR feed did not affect the stability of the bioreactors at both CLRs. The performance of the PADSBRs co-digesting swine carcasses was not statistically different from the control in terms of biogas production and quality. There was no accumulation of volatile fatty acids in the bioreactors at the end of the treatment cycle. The mixed-liquor pH and alkalinity remained within acceptable ranges for the anaerobic microflora. Also, there was no operational problem caused by the formation of foam and scum in the system.     

Psychrophilic anaerobic digestion of guinea pig manure in low-cost tubular digesters at high altitude

Guinea pig is one of the most common livestock in rural communities of the Andes. The aim of this research was to study the anaerobic digestion of guinea pig manure in low-cost unheated tubular digesters at high altitude. To this end, the performance of two pilot digesters was monitored during 7 months; and two greenhouse designs were compared. In the dome roof digester the temperature and biogas production were significantly higher than in the shed roof digester. However, the biogas production rate was low (0.04 m(biogas)(3)m(digester)(-3) d(-1)), which is attributed to the low organic loading rate (0.6 kg(VS)m(digester)(-3)d(-1)) and temperature (23°C) of the system, among other factors. In a preliminary fertilization study, the potato yield per hectare was increased by 100% using the effluent as biofertilizer. Improving manure management techniques, increasing the organic loading rate and co digesting other substrates may be considered to enhance the process.     

Thermophilic anaerobic digestion of source-sorted organic fraction of municipal solid waste

The influence of different organic fraction of municipal solid wastes during anaerobic thermophilic (55 degrees C) treatment of organic matter was studied in this work: food waste (FW), organic fraction of municipal solid waste (OFMSW) and shredded OFMSW (SH_OFMSW). All digester operated at dry conditions (20% total solids content) and were inoculated with 30% (in volume) of mesophilic digested sludge. Experimental results showed important different behaviours patterns in these wastes related with the organic matter biodegradation and biogas and methane production. The FW reactor showed the smallest waste biodegradation (32.4% VS removal) with high methane production (0.18 LCH4/gVS); in contrast the SH_OFMSW showed higher waste biodegradation (73.7% VS removal) with small methane production (0.05 LCH4/g VS). Finally, OFMSW showed the highest VS removal (79.5%) and the methane yield reached 0.08 LCH4/g VS. Therefore, the nature of organic substrate has an important influence on the biodegradation process and methane yield. Pre-treatment of waste is not necessary for OFMSW.     

Anaerobic Co-digestion of Swine Manure and Microalgae <Emphasis Type="Italic">Chlorella</Emphasis> sp.: Experimental Studies and Energy Analysis

Integration of algae production with livestock waste management has the potential to recover energy and nutrients from animal manure, while reducing discharges of organic matter, pathogens, and nutrients to the environment. In this study, microalgae Chlorella sp. were grown on centrate from anaerobically digested swine manure. The algae were harvested for mesophilic anaerobic digestion (AD) with swine manure for bioenergy production. Low biogas yields were observed in batch AD studies with algae alone, or when algae were co-digested with swine manure at ≥43 % algae (based on volatile solids [VS]). However, co-digestion of 6–16 % algae with swine manure produced similar biogas yields as digestion of swine manure alone. An average methane yield of 190 mL/g VS_fed was achieved in long-term semi-continuous co-digestion studies with 10?±?3 % algae with swine manure. Data from the experimental studies were used in an energy analysis assuming the process was scaled up to a concentrated animal feeding operation (CAFO) with 7000 pigs with integrated algae-based treatment of centrate and co-digestion of manure and the harvested algae. The average net energy production for the system was estimated at 1027 kWh per day. A mass balance indicated that 58 % of nitrogen (N) and 98 % of phosphorus (P) in the system were removed in the biosolids. A major advantage of the proposed process is the reduction in nutrient discharges compared with AD of swine waste without algae production.     

Anaerobic digestion of secondary residuals from an anaerobic bioreactor at a brewery to enhance bioenergy generation

Many beer breweries use high-rate anaerobic digestion (AD) systems to treat their soluble high-strength wastewater. Biogas from these AD systems is used to offset nonrenewable energy utilization in the brewery. With increasing nonrenewable energy costs, interest has mounted to also digest secondary residuals from the high-rate digester effluent, which consists of yeast cells, bacteria, methanogens, and small (hemi)cellulosic particles. Mesophilic (37 °C) and thermophilic (55 °C) lab-scale, low-rate continuously-stirred anaerobic digestion (CSAD) bioreactors were operated for 258 days by feeding secondary residuals at a volatile solids (VS) concentration of ∼40 g l⁻¹. At a hydraulic retention time (HRT) of 15 days and a VS loading rate of 2.7 g VS l⁻¹ day⁻¹, the mesophilic bioreactor showed an average specific volumetric biogas production rate of 0.88 l CH₄ l⁻¹ day⁻¹ and an effluent VS concentration of 22.2 g VS l⁻¹ (43.0% VS removal efficiency) while the thermophilic bioreactor displayed similar performances. The overall methane yield for both systems was 0.21 l CH₄ g⁻¹ VS fed and 0.47–0.48 l CH₄ g⁻¹ VS removed. A primary limitation of thermophilic digestion of this protein-rich waste is the inhibition of methanogens due to higher nondissociated (free) ammonia (NH₃) concentrations under similar total ammonium (NH₄ ⁺) concentrations at equilibrium. Since thermophilic AD did not result in advantageous methane production rates or yields, mesophilic AD was, therefore, superior in treating secondary residuals from high-rate AD effluent. An additional digester to convert secondary residuals to methane may increase the total biogas generation at the brewery by 8% compared to just conventional high-rate digestion of brewery wastewater alone. JIMB-2008: BioEnergy—Special issue.     

Influence of methane enrichment by aeration of recirculated supernatant on microbial activities during anaerobic digestion

A methane enrichment process (MEP) was evaluated that involved air purging of recycled digester contents to strip CO₂ and increase biogas methane content. The objective of this work was to determine if the aeration resulted in oxygen inhibition of microbial activities involved in anaerobic digestion of municipal solid waste. To assess the degree of biological perturbation associated with the MEP, the reactor effluent was sampled twice while the MEP was operating and twice while it was not operating. Analyses were run on composite samples (representing several different sample ports of the non-mixed reactor) and samples of digester effluent entering and leaving the MEP process. The analyses included volatile organic acids (VOA), effluent solids content, dehydrogenase activity, specific methanogenic activities (SMA), and enzyme-linked immunosorbent assay (ELISA) for methanogenic, sulfate-reducing, and cellulolytic bacterial species. Methane enrichment in these experiments occurred with the methane content of the biogas exceeding 90%. There were no effects of the MEP on effluent VOA concentration. The MEP had no effect on volatile solids (VS) levels of composite samples representing the non-mixed digester contents, however, VS was reduced in effluent passing through the MEP. Although MEP had an inhibitory effect on anaerobic populations leaving the MEP process, no inhibitory effects were observed in measurements of microbial activities in digester samples, including specific methane production rate, dehydrogenase, and numbers of specific organisms estimated using ELISA techniques.     

Inhibition of methanogenesis and its reversal during biogas formation from cattle manure

The composition of volatile fatty acids in the biogas digester based on cattle manure as substrate and stabilised at 25°C showed that it contained 87–88% branched chain fatty acids, comprising of isobutyric and isovaleric acids, in comparison to 38 % observed in the digester operating at 35°C. Mixed cellulolytic cultures equilibrated at 25°C (C-25) and 35‡C (C-35) showed similar properties, but rates of hydrolysis were three times higher than that observed in a standard biogas digester. The proportion of isobutyric and isovaleric were drastically reduced when C-25 was grown with glucose or filter paper as substrates. The volatile fatty acids recovered from C-25 (at 25°C) inhibited growth of methanogens on acetate, whereas that from C-35 was not inhibitory. The inhibitory effects were due to the branched chain fatty acids and were observed with isobutyric acid at concentrations as low as 50 ppm. Addition of another micro-organismRhodotorula selected for growth on isobutyric completely reversed this inhibition. Results indicate that the aceticlastic methanogens are very sensitive to inhibition by branched chain fatty acids and reduction in methane formation in biogas digester at lower temperature may be due to this effect.     

Effects of thermo-chemical pre-treatment of grass silage on methane production by anaerobic digestion

Dried grass silage (GS) was pre-treated at different NaOH loading rates (1%, 2.5%, 5% and 7.5% by volatile solids (VS) mass in grass silage) and temperatures (20 °C, 60 °C, 100 °C and 150 °C) to determine effects on its bio-degradability in terms of the hydrolysis yield and degradation of ligno-cellulosic materials for biogas production. At 100 °C and the four NaOH loadings, up to 45% of the total COD was solubilised and up to 65.6%, 36.1% and 21.2% of lignin, hemicellulose and cellulose were removed, respectively; biological methane production potentials obtained were 359.5, 401.8, 449.5 and 452.5 ml CH?/g VS added, respectively, being improved by 10-38.9% in comparison with untreated GS. VS removals following anaerobic digestion were 67.6%, 76.9%, 85.3%, 95.2% and 96.7% for untreated GS and GS treated at the four NaOH loadings, respectively. 100 °C and the NaOH loading rate of 5% is recommended as a proper GS pre-treatment condition.