Microbiological aspects of anaerobic digestion of swine slurry in upflow fixed-bed digesters with different packing materials

Samples taken from liquid influents, liquid effluents, bottom sediments and biofilms attached to the supports of three laboratory anaerobic, fixed-bed, upflow digesters, filled with wood chips, PVC or expanded-clay support media and fed with swine slurry, were tested microbiologically. The numbers of anaerobic heterotrophic, anaerobic cellulolytic, acidogenic-peptone-glucose fermenting and methanogenic bacteria were determined. For each digester the biogas production was monitored.The highest biogas production, referred to the volatile solids concentration in the feed, was obtained in the digester with wood chips, while production was almost nil in the digester with expanded clay.     

对处理不同废水的几种厌氧消化器生物量中优势产甲烷菌的观察

The biomass taken from AF, ABR, and UASB digesters treating municipal wastewater, swine waste and molassese stillage wastewater, municipal and molassese stillage wastewater respectively, were observed by light, epifluorescence microscope and scanning electron microscope. The prevalent methanogens in these digesters were Methanothrix. Exception for ABR carried out to treat swine waste, Methanosarcina with three forms of aggregations, cysts, and granules were presented in other digesters. A suggestion can be made that reactors with packing materials, such as, anaerobic filters, anaerobic contact beds and two-phase methanation digesters can be used to accumulate Methanosarcina due to their morphological, mu max, and Ks characteristics, in order to increase the wastewater treatment efficiency further.     

Vermicomposting of different forms of water hyacinth by the earthworm Eudrilus eugeniae,Kinberg

Six-month long trials were conducted on different vermireactors fed with one of the following forms of water hyacinth: (a) fresh whole plants, (b) dried whole plants, (c) chopped pieces of fresh plants, (d) 'spent' weed taken from reactors after extracting volatile fatty acids (VFAs), (e) precomposted fresh weed and (f) precomposted spent weed. The first four forms were studied with and without cowdung. The experiments revealed three clear trends (i) of the various forms of the weed assessed, the precomposted forms were the most favoured as feed by Eudrilus eugeniae, Kinberg, while the fresh whole form was the least favoured, (ii) the different forms of spent weed were favoured over the corresponding forms of fresh weed, and (iii) blending of cowdung (approximately 14% of the feed mass) with different forms of water hyacinth had a significant positive impact on vermicast output, growth in worm zoomass, and production of offspring relative to the corresponding unblended feed. In all reactors, the 'parent' earthworms steadily grew in size over the six-month span, and produced offspring. There was no mortality. The experiments thus confirm that water hyacinth can be sustainably vermicomposted in any of the forms with E. eugeniae.     

Improvement of activated sludge stabilisation and filterability during anaerobic digestion by fruit and vegetable waste addition

Anaerobic co-digestion of fruit and vegetable waste (FVW) and activated sludge (AS) was investigated using anaerobic sequencing batch reactors (ASBRs). The effects of AS:FVW ratio and the organic loading rate (OLR) on digesters performances were examined. The mixtures having AS:FVW ratios of 100:00, 65:35, 35:65, by a total solid (TS) basis were operated at an hydraulic retention time (HRT) of 20d. However, 30:70, 20:80, 15:85, 10:90 and 0:100 ratios were tested at an HRT of 10d. To investigate effects of aerobic and anaerobic digestion on the sludge filterability, specific resistance to filtration (R) was also determined. Increasing FVW proportions in the feedstock significantly improved the biogas production yield. The reactor that was fed with a 30:70 ratio showed the highest VS removal and biogas production yield of 88% and 0.57 L g(-1) VS added, respectively. The filterability results showed that the anaerobic effluent was characterised by a slightly better filterability efficiency of 1.6 x 10(16) m kg(-1) than 1.74 x 10(16) m kg(-1) of aerobic effluent. However, FVW addition improved the anaerobic co-digestion effluent filterability (5.52 x 10(14) m kg(-1)).     

Mesophilic and thermophilic anaerobic co-digestion of abattoir wastewater and fruit and vegetable waste in anaerobic sequencing batch reactors

Anaerobic co-digestion of fruit and vegetable waste (FVW) and abattoir wastewater (AW) was investigated using anaerobic sequencing batch reactors (ASBRs). The effects of hydraulic retention time (HRT) and temperature variations on digesters performances were examined. At both 20 and 10 days biogas production for co-digestion was greater thanks to the improved balance of nutrients. The high specific gas productions for the different digestion processes were 0.56, 0.61 and 0.85 l g⁻¹ total volatile solids (TVS) removal for digesters treating AW, FVW and AW + FVW, respectively. At an HRT of 20 days, biogas production rates from thermophilic digesters were higher on average than from mesophilic AW, FVW and AW + FVW digestion by 28.5, 44.5 and 25%, respectively. However, at 10 days of HRT results showed a decrease of biogas production rate for AW and AW + FVW digestion processes due to the high amount of free ammonia at high organic loading rate (OLR).     

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.     

Anaerobic digestion of municipal solid waste as a treatment prior to landfill

Anaerobic digestion of organic fraction of municipal solid waste was conducted in pilot-scale reactor based on high-solid combined anaerobic digestion process. This study was performed in two runs. In Run 1 and Run 2, pre-stage flushing and micro-aeration were conducted to determine their effect in terms of enhancing hydrolysis and acidification in ambient condition. In Run 2, after pre-stage, the methane phase (methanogenesis) was started-up after pH adjustment and inoculum addition in mesophilic condition. Acidified leachate produced in pre-stage was used for percolation during active methane phase. At the end of methane phase, air flushing was conducted before unloading the digesters. Hydrolysis and acidification yield of 140 g C/kg TS and 180 g VFA/kg TS were achieved, respectively in pre-stage. Micro-aeration exhibited an equivocal result in terms of enhancing hydrolysis/acidification; however it showed a positive effect in methane phase performance and this needed further investigation. Leachate percolation during methane phase showed an enhanced methanization when compared to the reactors without leachate percolation. After 60 days, 260 l CH(4)/kg VS was obtained. Based on the waste methane potential, 75% biogas conversion and 61% VS degradation were achieved.     

Anaerobic digestion of animal waste: effect of mixing

Six laboratory scale biogas mixed anaerobic digesters were operated to study the effect of biogas recycling rates and draft tube height on their performance. The digesters produced methane at 0.40-0.45 L per liter of digester volume per day. A higher methane production rate was observed in unmixed digesters, while increased biogas circulation rate reduced methane production. However, different draft tube heights caused no difference in the methane production rate. Air infiltration (up to 15% oxygen in the biogas) was observed in the digesters mixed by biogas recirculation. Slight air permeability of tubing or leakage on the vacuum side of the air pump may have caused the observed air infiltration. The similar performance of the mixed and unmixed digesters might be the result of the low solids concentration (50 g dry solids per liter of slurry) in the fed animal slurry, which could be sufficiently mixed by the naturally produced biogas.     

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.     

Methanogenic population dynamics during start-up of anaerobic digesters treating municipal solid waste and biosolids

An aggressive start-up strategy was used to initiate codigestion in two anaerobic, continuously mixed bench-top reactors at mesophilic (37 degrees C) and thermophilic (55 degrees C) conditions. The digesters were inoculated with mesophilic anaerobic sewage sludge and cattle manure and were fed a mixture of simulated municipal solid waste and biosolids in proportions that reflect U.S. production rates. The design organic loading rate was 3.1 kg volatile solids/m3/day and the retention time was 20 days. Ribosomal RNA-targeted oligonucleotide probes were used to determine the methanogenic community structure in the inocula and the digesters. Chemical analyses were performed to evaluate digester performance. The aggressive start-up strategy was successful for the thermophilic reactor, despite the use of a mesophilic inoculum. After a short start-up period (20 days), stable performance was observed with high gas production rates (1.52 m3/m3/day), high levels of methane in the biogas (59%), and substantial volatile solids (54%) and cellulose (58%) removals. In contrast, the mesophilic digester did not respond favorably to the start-up method. The concentrations of volatile fatty acids increased dramatically and pH control was difficult. After several weeks of operation, the mesophilic digester became more stable, but propionate levels remained very high. Methanogenic population dynamics correlated well with performance measures. Large fluctuations were observed in methanogenic population levels during the start-up period as volatile fatty acids accumulated and were subsequently consumed. Methanosaeta species were the most abundant methanogens in the inoculum, but their levels decreased rapidly as acetate built up. The increase in acetate levels was paralleled by an increase in Methanosarcina species abundance (up to 11.6 and 4.8% of total ribosomal RNA consisted of Methanosarcina species ribosomal RNA in mesophilic and thermophilic digesters, respectively). Methanobacteriaceae were the most abundant hydrogenotrophic methanogens in both digesters, but their levels were higher in the thermophilic digester.     

Influence of the dimensional characteristics of polyurethane foam on high rate anaerobic digestion of piggery manure

Summay Completely mixed anaerobic reactors containing ca 10 % of their working volume as polyurethane (PUR) foam, can digest heterogenous slurries such as pig manure in a stable way at short hydraulic retention times of the order of 6.0 to 7.5 day. The influence of pore size and pad size of the PUR foam on the process was investigated. The PUR foam with the smallest pore size investigated (i.e. 45 pores per inch) gave the best biogas yield and biogas production rate. Pad size seemed to be of minor importance, having no effect on the performance of the digesters within the range examined (1 to 3.6 cm rib length). Apparently, diffusion of fatty acids into the PUR pads appeared not to be rate limiting under the lab scale reactor conditions examined.     

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.     

Weed Seed Survival during Anaerobic Digestion in Biogas Plants

Anaerobic digestion using animal manure and crop biomass is increasingly being used to produce biogas as a durable alternative to fossil fuel. The sludge, the leftover after processing, is returned to the field as a crop fertilizer. If weed seeds survive anaerobic digestion, the use of contaminated sludge poses a phytosanitary risk. The conditions that seeds are likely to encounter in biogas plants, and the effect of these, in particular temperature, on seed viability were reviewed. Knowledge on seed defence mechanisms and how these might protect seeds from inactivation in biogas reactors was summarized. Mechanisms of seed inactivation can be classified as thermal, biological and chemical. Weed species with hard seeds (physical dormant), high thermoresistance, a thick seed coat or adapted to endozoochory were identified as high-risk species. Specific seed traits could be used in future tests to circumvent extensive testing of seeds in biogas reactors.     

Inhibition of Biogas Production by Alkyl Benzene Sulfonates (LAS) in a Screening Test for Anaerobic Biodegradability

The effect of the inoculum source on the digestion of linear alkylbenzene sulfonates (LAS) under anaerobic conditions has been investigated. The potential for primary and ultimate LAS biodegradation of anaerobic sludge samples obtained from wastewater treatment plants (WWTPs) of different geographical locations was studied applying a batch test system. It was found that only 4–22% of the LAS added to the batch anaerobic digesters was primarily transformed suggesting a poor primary degradation of the LAS molecule in anaerobic discontinuous systems. Regarding ultimate biodegradation, the addition of LAS to the batch anaerobic digesters caused a reduction on the extent of biogas production. Significant differences in the inhibition extent of the biogas production were observed (4–26%) depending on the sludge used as inoculum. Effect of the surfactant on the anaerobic microorganisms was correlated with its concentration in the aqueous phase. Sorption of LAS on anaerobic sludge affects its toxicity by depletion of the available fraction of the surfactant. LAS content on sludge was related to the total amount of calcium and magnesium extractable ions. The presence of divalent cations promote the association of LAS with anaerobic sludge reducing its bioavailability and the extent of its inhibitory effect on the biogas production.     

Characteristics and biogas production potential of municipal solid wastes pretreated with a rotary drum reactor

This study was conducted to determine the characteristics and biogas production potential of organic materials separated from municipal solid wastes using a rotary drum reactor (RDR) process. Four different types of wastes were first pretreated with a commercial RDR system at different retention times (1, 2 and 3 d) and the organic fractions were tested with batch anaerobic digesters with 2.6 g VS L(-1) initial loading. The four types of waste were: municipal solid waste (MSW), a mixture of MSW and paper waste, a mixture of MSW and biosolids, and a mixture of paper and biosolids. After 20 d of thermophilic digestion (50+/-1 degrees C), it was found that the biogas yields of the above materials were in the range of 457-557 mL g VS(-1) and the biogas contained 57.3-60.6% methane. The total solid and volatile solid reductions ranged from 50.2% to 65.0% and 51.8% to 66.8%, respectively. For each material, the change of retention time in the RDR from 1 to 3d did not show significant (alpha=0.05) influence on the biogas yields of the recovered organic materials. Further studies are needed to determine the minimum retention time requirements in the RDR system to achieve effective separation of organic from inorganic materials and produce suitable feedstock for anaerobic digesters.     

Effect of temperature on the performance of an anaerobic tubular reactor treating fruit and vegetable waste

This study compares the performance of anaerobic digestion of fruit and vegetable waste (FVW) in the thermophilic (55 °C) process with those under psychrophilic (20 °C) and mesophilic (35 °C) conditions in a tubular anaerobic digesters on a laboratory scale. The hydraulic retention time (HRT) ranged from 10 to 20 days, and raw fruit and vegetable waste was supplied in a semi-continuous mode at various concentrations of total solids (TS) (4, 6, 8 and 10% on dry weight). Biogas production from the experimental thermophilic digester was higher on average than from psychrophilic and mesophilic digesters by 144 and 41%, respectively. The net energy production in the thermophilic digester was 195.7 and 49.07 kJ per day higher than that for the psychrophilic and mesophilic digesters, respectively. The relation between the daily production of biogas and the temperature indicates that for the same produced quantity of biogas, the size of the thermophilic digester can be reduced with regard to that of the psychrophilic and the mesophilic digesters.     

Treatment of landfill leachate using UASB-MBR-SHARON–Anammox configuration

Leachate treatment is a challenging issue due to its high pollutant loads. There are several studies on feasible treatment methods of leachate. In the scope of this study, high organic content of young leachate was eliminated using an upflow anaerobic sludge blanket (UASB) and a membrane bioreactor (MBR) in sequence and effluent of the system was given to single reactor for high activity ammonia removal over nitrite (SHARON) and anaerobic ammonia oxidation (Anammox) reactors to remove nitrogen content. All reactors were set up at lab scale in order to evaluate the usage of these processes in sequencing order for leachate treatment. COD and TKN removal efficiencies were over 90 % in the combined processes which were operated during the study. The biodegradable portion of organic matter was removed with an efficiency of 99 %. BOD₅ concentration decreased to 50 mg/L by UASB and MBR in sequence even the influent BOD₅ concentration was over 8,000 mg/L. Although high nitrogen concentrations were observed in raw leachate, successful removal of nitrogen was accomplished by consecutive operations of SHARON and Anammox reactors. The results of this study demonstrated that with an efficient pretreatment of leachate, the combination of SHARON–Anammox processes is an effective method for the treatment of high nitrogen content in leachate.     

Enrichment and description of novel bacteria performing syntrophic propionate oxidation at high ammonia level

Inefficient syntrophic propionate degradation causes severe operating disturbances and reduces biogas productivity in many high-ammonia anaerobic digesters, but propionate-degrading microorganisms in these systems remain unknown. Here, we identified candidate ammonia-tolerant syntrophic propionate-oxidising bacteria using propionate enrichment at high ammonia levels (0.7–0.8 g NH₃ L⁻¹) in continuously-fed reactors. We reconstructed 30 high-quality metagenome-assembled genomes (MAGs) from the propionate-fed reactors, which revealed two novel species from the families Peptococcaceae and Desulfobulbaceae as syntrophic propionate-oxidising candidates. Both MAGs possess genomic potential for the propionate oxidation and electron transfer required for syntrophic energy conservation and, similar to ammonia-tolerant acetate degrading syntrophs, both MAGs contain genes predicted to link to ammonia and pH tolerance. Based on relative abundance, a Peptococcaceae sp. appeared to be the main propionate degrader and has been given the provisional name “Candidatus Syntrophopropionicum ammoniitolerans”. This bacterium was also found in high-ammonia biogas digesters, using quantitative PCR. Acetate was degraded by syntrophic acetate-oxidising bacteria and the hydrogenotrophic methanogenic community consisted of Methanoculleus bourgensis and a yet to be characterised Methanoculleus sp. This work provides knowledge of cooperating syntrophic species in high-ammonia systems and reveals that ammonia-tolerant syntrophic propionate-degrading populations share common features, but diverge genomically and taxonomically from known species.     

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.     

High-rate composting-vermicomposting of water hyacinth (Eichhornia crassipes,Mart. Solms)

In an attempt to develop a system with which the aquatic weed water hyacinth (Eichhornia crassipes, Mart. Solms) can be economically processed to generate vermicompost in large quantities, the weed was first composted by a 'high-rate' method and then subjected to vermicomposting in reactors operating at much larger densities of earthworm than recommended hitherto: 50, 62.5, 75, 87.5, 100, 112.5, 125, 137.5, and 150 adults of Eudrilus eugeniae Kinberg per litre of digester volume. The composting step was accomplished in 20 days and the composted weed was found to be vermicomposted three times as rapidly as uncomposted water hyacinth [Bioresource Technology 76 (2001) 177]. The studies substantiated the feasibility of high-rate composting-vermicomposting systems, as all reactors yielded consistent vermicast output during seven months of operation. There was no earthworm mortality during the first four months in spite of the high animal densities in the reactors. In the subsequent three months a total of 79 worms died out of 1650, representing less than 1.6% mortality per month. The results also indicated that an increase in the surface-to-volume ratio of the reactors might further improve their efficiency.