An innovative online VFA monitoring system for the anerobic process, based on headspace gas chromatography

A new method for online measurement of volatile fatty acids (VFA) in anerobic digesters has been developed based on headspace gas chromatography (HSGC). The method applies ex situ VFA stripping with variable headspace volume and gas analysis by gas chromatography-flame ionization detection (GC-FID). In each extraction, digester sample was acidified with H(3)PO(4) and NaHSO(4), then heated to strip the VFA into the gas phase. The gas was sampled in a low friction glass syringe before injected into the GC for measurement. The system has been tested for online monitoring of a lab-scale CSTR reactor treating manure for more than 6 months and has shown good agreement with off-line analysis. The system is capable of measuring individual VFA components. This is of advantage since specific VFA components such as propionic and butyric acid can give extra information about the process status. Another important advantage of this sensor is that there is no filtration, which makes possible application in high solids environments. The system can thus be easily applied in a full-scale biogas reactor by connecting the system to the liquid circulation loop to obtain fresh sample from the reactor. Local calibration is needed but automatic calibration is also possible using standard addition method. Sampling duration is 25-40 min, depending on the washing duration, and sensor response is 10 min. This is appropriate for full-scale reactors, since dynamics within most biogas reactors are of the order of several hours.     

Recovery of methane-rich gas from solid-feed anaerobic digestion of ipomoea (Ipomoea carnea)

Studies are presented on new types of anaerobic digesters in which chopped or dry crushed Ipomoea carnea was fed without any other pretreatment, in an attempt to develop commercially viable means of utilizing the otherwise very harmful plant. Two types of solid-feed anaerobic digesters (SFADs) were studied. The first type had a single vessel in which the bottom 35% portion was separated from the top portion by a perforated PVC disk. The weed was charged from the top and inoculated with anaerobically digested cowdung-water slurry. The fermentation of the weed in the reactor led to the formation of volatile fatty acids (VFAs) plus some biogas. The leachate, rich in the VFAs, was passed through the perforated PVC sheet and collected in the lower portion of the vessel. The other type of reactors had two vessels, the first one was fully charged with the weed and the second received the VFA leachate. With both types were attached upflow anaerobic filters (UAFs) which converted the leachate into combustible biogas consisting of approximately 70% methane. All SFADs developed very consistent performance in terms of biogas yield within 17 weeks of start. The two-compartment reactors yielded significantly more biogas than the single-compartment reactors of corresponding total volume, and the reactors with which anaerobic filters (AF) were attached yielded more biogas than the ones without AF. The best performing units generated 2.41m(3) of biogas per m(3) of digester volume, as compared to 0.1-0.2m(3) of biogas, m(-3)d(-1), obtainable with conventional digesters. This indicates the viability of this technology. The spent weed can be vermicomposted directly to obtain good soil-conditioner cum fertilizer; earthworm Eudrilus eugeniae produced 540mg vermicast per animal every day, achieving near total conversion of feed to vermicast in 20 days. The proposed systems, thus, makes it possible to accomplish total utilization of ipomoea.     

Dynamics of the anaerobic process: effects of volatile fatty acids

A complex and fast dynamic response of the anaerobic biogas system was observed when the system was subjected to pulses of volatile fatty acids (VFAs). It was shown that a pulse of specific VFAs into a well-functioning continuous stirred tank reactor (CSTR) system operating on cow manure affected both CH(4) yield, pH, and gas production and that a unique reaction pattern was seen for the higher VFAs as a result of these pulses. In this study, two thermophilic laboratory reactors were equipped with a novel VFA-sensor for monitoring specific VFAs online. Pulses of VFAs were shown to have a positive effect on process yield and the levels of all VFA were shown to stabilize at a lower level after the biomass had been subjected to several pulses. The response to pulses of propionate or acetate was different from the response to butyrate, iso-butyrate, valerate, or iso-valerate. High concentrations of propionate affected the degradation of all VFAs, while a pulse of acetate affected primarily the degradation of iso-valerate or 2-methylbutyrate. Pulses of n-butyrate, iso-butyrate, and iso-valerate yielded only acetate, while degradation of n-valerate gave both propionate and acetate. Product sensitivity or inhibition was shown for the degradation of all VFAs tested. Based on the results, it was concluded that measurements of all specific VFAs are important for control purposes and increase and decrease in a specific VFA should always be evaluated in close relationship to the conversion of other VFAs and the history of the reactor process. It should be pointed out that the observed dynamics of VFA responses were based on hourly measurements, meaning that the response duration was much lower than the hydraulic retention time, which exceeds several days in anaerobic CSTR systems.     

Influence of environmental conditions on methanogenic compositions in anaerobic biogas reactors

The influence of environmental parameters on the diversity of methanogenic communities in 15 full-scale biogas plants operating under different conditions with either manure or sludge as feedstock was studied. Fluorescence in situ hybridization was used to identify dominant methanogenic members of the Archaea in the reactor samples; enriched and pure cultures were used to support the in situ identification. Dominance could be identified by a positive response by more than 90% of the total members of the Archaea to a specific group- or order-level probe. There was a clear dichotomy between the manure digesters and the sludge digesters. The manure digesters contained high levels of ammonia and of volatile fatty acids (VFA) and were dominated by members of the Methanosarcinaceae, while the sludge digesters contained low levels of ammonia and of VFA and were dominated by members of the Methanosaetaceae. The methanogenic diversity was greater in reactors operating under mesophilic temperatures. The impact of the original inoculum used for the reactor start-up was also investigated by assessment of the present population in the reactor. The inoculum population appeared to have no influence on the eventual population.     

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.     

Potential of biogas recirculation to enhance biomass accumulation on supporting media

Two lab-scale anaerobic hybrid reactors (AHR) were operated to investigate the effect of recirculated biogas on the development of biomass on supporting media during the start-up. The reactor comprised of two distinct zones; sludge bed on the bottom and packed bed using nylon fiber as the media on the upper half of the reactor. Both reactors were continuously fed with cassava starch wastewater. The organic loading rate (OLR) was increased from 0.3 to 5.5 g COD/L/day by gradually decreasing the hydraulic retention time (HRT) from 37 to 3.5 days in two months. The biogas at 2.6 L/L/day was recirculated merely in the first month of the operation in order to allow the attached biomass to grow according to the organic matters present in the reactor at the final stage of the start up. Chemical oxygen demand (COD) removal efficiency of over 80% was achieved throughout the study. The result demonstrated a better COD removal efficiency for the reactor with biogas recirculation, especially at low HRTs. The amounts of biomass accumulated on the media in both reactors were slightly different with 11.9 gVSS found on the one with biogas recirculation compared to 9.8 gVSS on the other. In addition, 16.3% increase of the sludge bed was achieved with biogas recirculation as opposed to 9% in the control one. The attached biomass activity test indicated a greater amount and more favorable ratio of the methanogenic bacterial group on the media with the recirculation correlating well to a relatively higher methane content in biogas. As a result, the recirculation of biogas has a potential of improving the characteristics of the AHR especially in terms of biomass accumulation.     

Influence of Environmental Conditions on Methanogenic Compositions in Anaerobic Biogas Reactors

The influence of environmental parameters on the diversity of methanogenic communities in 15 full-scale biogas plants operating under different conditions with either manure or sludge as feedstock was studied. Fluorescence in situ hybridization was used to identify dominant methanogenic members of the Archaea in the reactor samples; enriched and pure cultures were used to support the in situ identification. Dominance could be identified by a positive response by more than 90% of the total members of the Archaea to a specific group- or order-level probe. There was a clear dichotomy between the manure digesters and the sludge digesters. The manure digesters contained high levels of ammonia and of volatile fatty acids (VFA) and were dominated by members of the Methanosarcinaceae, while the sludge digesters contained low levels of ammonia and of VFA and were dominated by members of the Methanosaetaceae. The methanogenic diversity was greater in reactors operating under mesophilic temperatures. The impact of the original inoculum used for the reactor start-up was also investigated by assessment of the present population in the reactor. The inoculum population appeared to have no influence on the eventual population.     

Volatile fatty acids (VFAs) and methane from food waste and cow slurry: Comparison of biogas and VFA fermentation processes

The potential of various biomasses for the production of green chemicals is currently one of the key topics in the field of the circular economy. Volatile fatty acids (VFAs) are intermediates in the methane formation pathway of anaerobic digestion and they can be produced in similar reactors as biogas to increase the productivity of a digestion plant, as VFAs have more varying end uses compared to biogas and methane. In this study, the aim was to assess the biogas and VFA production of food waste (FW) and cow slurry (CS) using the anaerobic biogas plant inoculum treating the corresponding substrates. The biogas and VFA production of both biomasses were studied in identical batch scale laboratory conditions while the process performance was assessed with chemical and microbial analyses. As a result, FW and CS were shown to have different chemical performances and microbial dynamics in both VFA and biogas processes. FW as a substrate showed higher yields in both processes (435 ml CH₄/g VS_fed and 434 mg VFA/g VS_fed) due to its characteristics (pH, organic composition, microbial communities), and thus, the vast volume of CS makes it also a relevant substrate for VFA and biogas production. In this study, VFA profiles were highly dependent on the substrate and inoculum characteristics, while orders Clostridiales and Lactobacillales were connected with high VFA and butyric acid production with FW as a substrate. In conclusion, anaerobic digestion supports the implementation of the waste management hierarchy as it enables the production of renewable green chemicals from both urban and rural waste materials.     

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.     

Comparison of two-stage thermophilic (68 degrees C/55 degrees C) anaerobic digestion with one-stage thermophilic (55 degrees C) digestion of cattle manure

A two-stage 68 degrees C/55 degrees C anaerobic degradation process for treatment of cattle manure was studied. In batch experiments, an increase of the specific methane yield, ranging from 24% to 56%, was obtained when cattle manure and its fractions (fibers and liquid) were pretreated at 68 degrees C for periods of 36, 108, and 168 h, and subsequently digested at 55 degrees C. In a lab-scale experiment, the performance of a two-stage reactor system, consisting of a digester operating at 68 degrees C with a hydraulic retention time (HRT) of 3 days, connected to a 55 degrees C reactor with 12-day HRT, was compared with a conventional single-stage reactor running at 55 degrees C with 15-days HRT. When an organic loading of 3 g volatile solids (VS) per liter per day was applied, the two-stage setup had a 6% to 8% higher specific methane yield and a 9% more effective VS-removal than the conventional single-stage reactor. The 68 degrees C reactor generated 7% to 9% of the total amount of methane of the two-stage system and maintained a volatile fatty acids (VFA) concentration of 4.0 to 4.4 g acetate per liter. Population size and activity of aceticlastic methanogens, syntrophic bacteria, and hydrolytic/fermentative bacteria were significantly lower in the 68 degrees C reactor than in the 55 degrees C reactors. The density levels of methanogens utilizing H2/CO2 or formate were, however, in the same range for all reactors, although the degradation of these substrates was significantly lower in the 68 degrees C reactor than in the 55 degrees C reactors. Temporal temperature gradient electrophoresis profiles (TTGE) of the 68 degrees C reactor demonstrated a stable bacterial community along with a less divergent community of archaeal species.     

Methanogenesis in thermophilic biogas reactors

Methanogenesis in thermophilic biogas reactors fed with different wastes is examined. The specific methanogenic activity with acetate or hydrogen as substrate reflected the organic loading of the specific reactor examined. Increasing the loading of thermophilic reactors stabilized the process as indicated by a lower concentration of volatile fatty acids in the effluent from the reactors. The specific methanogenic activity in a thermophilic pilot-plant biogas reactor fed with a mixture of cow and pig manure reflected the stability of the reactor. The numbers of methanogens counted by the most probable number (MPN) technique with acetate or hydrogen as substrate were further found to vary depending on the loading rate and the stability of the reactor. The numbers of methanogens counted with antibody probes in one of the reactor samples was 10 times lower for the hydrogen-utilizing methanogens compared to the counts using the MPN technique, indicating that other non-reacting methanogens were present. Methanogens that reacted with the probe againstMethanobacterium thermoautotrophicum were the most numerous in this reactor. For the acetate-utilizing methanogens, the numbers counted with the antibody probes were more than a factor of 10 higher than the numbers found by MPN. The majority of acetate utilizing methanogens in the reactor wereMethanosarcina spp. single cells, which is a difficult form of the organism to cultivatein vitro. No reactions were observed with antibody probes raised againstMethanothrix soehngenii orMethanothrix CALS-1 in any of the thermophilic biogas reactors examined. Studies using 2-¹⁴C-labeled acetate showed that at high concentrations (more than approx. 1 mM) acetate was metabolized via the aceticlastic pathway, transforming the methyl-group of acetate into methane. When the concentration of acetate was less than approx. 1 mM, most of the acetate was oxidized via a two-step mechanism (syntrophic acetate oxidation) involving one organism oxidizing acetate into hydrogen and carbon dioxide and a hydrogen-utilizing methanogen forming the products of the first microorganism into methane. In thermophilic biogas reactors, acetate oxidizing cultures occupied the niche ofMethanothrix species, aceticlastic methanogens which dominate at low acetate concentrations in mesophilic systems. Normally, thermophilic biogas reactors are operated at temperatures from 52 to 56° C. Experiments using biogas reactors fed with cow manure showed that the same biogas yield found at 55° C could be obtained at 61° C after a long adaptation period. However, propionate degradation was inhibited by increasing the temperature.     

分段组合式厌氧反应器预警性能

采用模拟有机废水,研究了分段组合式厌氧反应器 (Compartmentalized anaerobic reactor,CAR) 的预警性能。试验结果表明：高效厌氧反应器的运行稳定性低于常效厌氧反应器。在高效工况下,进水的化学需氧量 (Chemical oxygen demand,COD) 浓度的小幅提升 (平均相对标准偏差为8.08％) 可引起出水COD浓度 (平均相对标准偏差为32.95％) 和出水挥发性脂肪酸 (Volatile fatty acids,VFA) 浓度 (平均相对标准偏差为40.46％) 的大幅增加。容积负荷饱和度和VFA饱和度与反应器工况密切相关,可用于厌氧反应过程的预警。常负荷工况下 (容积负荷饱和度和VFA饱和度低于0.89与0.40),反应运行性能稳定;满负荷工况下 (容积负荷饱和度和VFA饱和度趋近1),反应运行性能波动增大;超负荷工况下 (容积负荷饱和度和VFA饱和度超过1),反应运行性能恶化。     

Evolution of microorganisms in thermophilic-dry anaerobic digestion

Microbial population dynamics were studied during the start-up and stabilization periods in thermophilic-dry anaerobic digestion at lab-scale. The experimental protocol was defined to quantify Eubacteria and Archaea using Fluorescent in situ hybridization (FISH) in a continuously stirred tank reactor (CSTR), without recycling solids. The reactor was subjected to a programme of steady-state operation over a range of the retention times from 40 to 25 days, with an organic loading rate between 4.42 and 7.50 kg volatile solid/m3/day. Changes in microbial concentrations were linked to traditional performance parameters such as biogas production and VS removal. The relations of Eubacteria:Archaea and H2-utilising methanogens:acetate-utilising methanogens were 88:12 and 11:1, respectively, during start-up stage. Hydrogenotrophic methanogens, although important in the initial phase of the reactor start-up, were displaced by acetoclastic methanogens at steady-state, thus their relation were 7:32, respectively. The methane yield coefficient, the methane content in the biogas and VS removal were stabilized around 0.30 LCH4/gCOD, 50% and 80%, respectively. Methanogenic population correlated well with performance measurements.     

Microbial populations associated with fixed- and floating-bed reactors during a two-stage anaerobic process

Microbial populations associated with methanogenic fixed- or floating-bed bioreactors used for anaerobic digestion of lignocellulosic waste were investigated. Fluorescent in situ hybridization (FISH) was used to characterize microorganisms in samples obtained from different heights in the reactors, which were operated in a semi-continuous manner (feeding and mixing once every 2 days). The FISH results showed that Methanosaeta concilii cells were most numerous at the bottom of both reactors. M. concilii cells were more abundant in the fixed-bed reactor (FXBR), which performed better than the floating-bed reactor (FLBR). Species of the Methanosarcina genera (mainly M. barkeri and M. mazei) were also observed in the FLBR but rarely in the FXBR. Methane production in each of the reactors ranged from 0.29 to 0.33 m3 CH(4)/kg COD(rem) (chemical oxygen demand removed). The removal of volatile fatty acids (VFA; 70-75 h) in the FXBR was more efficient than in the FLBR.     

Anaerobic treatment of winery wastewater using laboratory-scale multi- and single-fed filters at ambient temperatures

A lab-scale investigation was conducted to examine the effectiveness of a multi-fed upflow anaerobic filter process for the methane production from a rice winery effluent at ambient temperatures. The experiment was carried in two identical 3.0-l upflow filters, a single-fed reactor and a multi-fed reactor. The results showed that the multi-fed reactor, operated at the ambient temperatures of 19–27 °C and influent chemical oxygen demand (COD) of 8.34–25.76 g/l, could remove over 82% of COD even at an organic loading rate (OLR) of 37.68 g-COD/l d and a short hydraulic retention time (HRT) of 8 h. This reactor produced biogas with a methane yield of 0.30–0.35 l-CH₄/g-COD_removed. The multi-fed upflow anaerobic filter was proved to be more efficient than the single-fed reactor in terms of COD removal efficiency and stability against hydraulic loading shocks. A linear-regression model with influent COD concentration and HRT terms adequately described the multi-fed upflow anaerobic filter system for the treatment of rice winery wastewater at ambient temperatures.     

Anaerobic Codigestion of Sludge: Addition of Butcher’s Fat Waste as a Cosubstrate for Increasing Biogas Production

Fat waste discarded from butcheries was used as a cosubstrate in the anaerobic codigestion of sewage sludge (SS). The process was evaluated under mesophilic and thermophilic conditions. The codigestion was successfully attained despite some inhibitory stages initially present that had their origin in the accumulation of volatile fatty acids (VFA) and adsorption of long-chain fatty acids (LCFA). The addition of a fat waste improved digestion stability and increased biogas yields thanks to the higher organic loading rate (OLR) applied to the reactors. However, thermophilic digestion was characterized by an effluent of poor quality and high VFA content. Results from spectroscopic analysis suggested the adsorption of lipid components onto the anaerobic biomass, thus disturbing the complete degradation of substrate during the treatment. The formation of fatty aggregates in the thermophilic reactor prevented process failure by avoiding the exposure of biomass to the toxic effect of high LCFA concentrations.     

Different types of sludge granules in UASB reactors treating acidified wastewaters

The influence of a high energy substrate, i.e. sucrose, on the granular sludge yield and the development of different types of granular sludge was investigated by using Upflow Anaerobic Sludge Bed (UASB) reactors fed with synthetic wastewater. The feed COD was a mixture of volatile fatty acids (VFA) i.e., 20, 40, and 40% of the COD as C₂-, C₃-, and C₄-VFA, respectively. Furthermore, experiments were carried out in which 10 and 30% of the VFA COD was substituted with sucrose. The following distinctly different types of granules were observed in each testrun: in the reactor fed with solely VFA, black (B) and white (W) granules developed; in the reactor fed with a mixture of 90% VFA and 10% sucrose, three types of granules i.e., B, W, and grey (G) granules could be seen; in the reactor fed with 70% VFA and 30% sucrose, only W and G granules were found. The granular sludge yield increased proportional to the amount of sucrose COD. At steady-state performance of the reactors, specific acidogenic (SAA) and methanogenic (SMA) activity tests on these granules revealed that B granules had the highest SMA with low SAA. The W granules had very high SMA with low SAA. G granules gave the highest SAA with a considerable SMA. Measurement of coenzyme F₄₂₀ revealed that B granules consist mainly of acetoclastic methanogens. The fore-mentioned tests were supplemented with analyses of the wash-out cells present in the reactor effluent and the results suggested that acidogens, if present, prevail at the granule surface. The B granules were particularly rich in Ca, Mn, and Zn minerals. The size distribution analysis showed that the granule diameter increased in the following order: B相似文献   

Microbial communities involved in biogas production from wheat straw as the sole substrate within a two-phase solid-state anaerobic digestion

Microbial communities involved in biogas production from wheat straw as the sole substrate were investigated. Anaerobic digestion was carried out within an up-flow anaerobic solid-state (UASS) reactor connected to an anaerobic filter (AF) by liquor recirculation. Two lab-scale reactor systems were operated simultaneously at 37 °C and 55 °C. The UASS reactors were fed at a fixed organic loading rate of 2.5 g L⁻¹ d⁻¹, based on volatile solids. Molecular genetic analyses of the bacterial and archaeal communities within the UASS reactors (digestate and effluent liquor) and the AFs (biofilm carrier and effluent liquor) were conducted under steady-state conditions. The thermophilic UASS reactor had a considerably higher biogas and methane yield in comparison to the mesophilic UASS, while the mesophilic AF was slightly more productive than the thermophilic AF. When the thermophilic and mesophilic community structures were compared, the thermophilic system was characterized by a higher Firmicutes to Bacteroidetes ratio, as revealed by 16S rRNA gene (rrs) sequence analysis. The composition of the archaeal communities was phase-separated under thermophilic conditions, but rather stage-specific under mesophilic conditions. Family- and order-specific real-time PCR of methanogenic Archaea supported the taxonomic distribution obtained by rrs sequence analysis. The higher anaerobic digestion efficiency of the thermophilic compared to the mesophilic UASS reactor was accompanied by a high abundance of Firmicutes and Methanosarcina sp. in the thermophilic UASS biofilm.     

Effect of mixing on biogas production during mesophilic anaerobic digestion of screened dairy manure in a pilot plant

The effect of mixing on biogas production of a 1.5‐m³ pilot continuous stirred tank reactor (CSTR) processing screened dairy manure was evaluated. Mixing was carried out by recirculation of reactor content with a mono pump. The experiment was conducted at a controlled temperature of 37±1°C and hydraulic retention times (HRTs) of 20 and 10 days. The effect of continuous and intermittent operation of the recirculation pump on biogas production was studied. At 10 days of HRT, the results showed a minimal influence of recirculation rate on biogas production and that continuous recirculation did not improve reactor performance. At 20 days of HRT, the recirculation rate did not affect reactor performance. Combination of low solid content in feed animal slurry and long HRTs results in minimal mixing requirements for anaerobic digestion.