Oxygen tolerance capacity of upflow anaerobic solid-state (UASS) with anaerobic filter (AF) system.

In order to investigate the oxygen tolerance capacity of upflow anaerobic solid-state (UASS) with anaerobic filter (AF) system, the effect of microaeration on thermophilic anaerobic digestion of maize straw was investigated under batch conditions and in the UASS with AF system. Aeration intensities of 0-431mL O2/gvs were conducted as pretreatment under batch conditions. Aeration pretreatment obviously enhanced anaerobic digestion and an aeration intensity of 431mL O2/gvs increased the methane yield by 82.2%. Aeration intensities of 0-355mL O2/gvs were conducted in the process liquor circulation of the UASS with AF system. Dissolved oxygen (DO) of UASS and AF reactors kept around 1.39±0.27 and 0.99±0.38mg/L, respectively. pH was relatively stable around 7.11±0.04. Volatile fatty acids and soluble chemical oxygen demand concentration in UASS reactor were higher than those in AF reactor. Methane yield of the whole system was almost stable at 85±7mL/gvs as aeration intensity increased step by step. The UASS with AF system showed good oxygen tolerance capacity.

Effect of hydrothermally carbonized char application on trace gas emissions from two sandy soil horizons.

The application of biochar to soil is a potential tool for the long-term sequestration of C and a possible mitigation of greenhouse gas (GHG) emissions. Among the various processes available to produce biochar, hydrothermal carbonization is one technique that is suitable for moist feedstock like digestates from biogas production. The aim of this study was to investigate the stability of C and emissions of NO after the addition of (i) digested wheat ( L.) straw (digestate) and (ii) hydrothermally carbonized (HTC) char of wheat straw as well as (iii) HTC char of digested wheat straw to two soil horizons that differed in C content. The HTC chars were obtained from wheat straw and digested wheat straw that were hydrothermally carbonized at 230°C for 6 h. The digestate and HTC chars were mixed with soil and incubated in 125-mL vessels. The GHG emissions of CO and NO were measured at regular intervals. Additionally, after 108 d, N was applied in the form of NHNO equivalent to 100 kg N ha. After 500 d of incubation, the digestate had lost 34% of C, while the soil mixture with the corresponding HTC char lost 12% of C in the form of CO from the topsoil. The estimated bi-exponential half-life of the recalcitrant C was more than 50% longer for the carbonized material than for the untreated digestate. The NO emissions from both HTC chars were significantly reduced compared with untreated digestate. The reductions were up to 64% for the topsoil and 60% for the subsoil samples. These laboratory results show that HTC holds the potential to increase the C stability of fermented and carbonized biomasses and to reduce NO emissions.

Development of a flow-fluorescence in situ hybridization protocol for the analysis of microbial communities in anaerobic fermentation liquor.

BACKGROUND: The production of bio-methane from renewable raw material is of high interest because of the increasing scarcity of fossil fuels. The process of biomethanation is based on the inter- and intraspecific metabolic activity of a highly diverse and dynamic microbial community. The community structure of the microbial biocenosis varies between different biogas reactors and the knowledge about these microbial communities is still fragmentary. However, up to now no approaches are available allowing a fast and reliable access to the microbial community structure. Hence, the aim of this study was to originate a Flow-FISH protocol, namely a combination of flow cytometry and fluorescence in situ hybridization, for the analysis of the metabolically active microorganisms in biogas reactor samples. With respect to the heterogenic texture of biogas reactor samples and to collect all cells including those of cell aggregates and biofilms the development of a preceding purification procedure was indispensable. RESULTS: Six different purification procedures with in total 29 modifications were tested. The optimized purification procedure combines the use of the detergent sodium hexametaphosphate with ultrasonic treatment and a final filtration step. By this treatment, the detachment of microbial cells from particles as well as the disbandment of cell aggregates was obtained at minimized cell loss. A Flow-FISH protocol was developed avoiding dehydration and minimizing centrifugation steps. In the exemplary application of this protocol on pure cultures as well as biogas reactor samples high hybridization rates were achieved for commonly established domain specific oligonucleotide probes enabling the specific detection of metabolically active bacteria and archaea. Cross hybridization and autofluorescence effects could be excluded by the use of a nonsense probe and negative controls, respectively. CONCLUSIONS: The approach described in this study enables for the first time the analysis of the metabolically active fraction of the microbial communities within biogas reactors by Flow-FISH.

Characterization of the methanogenic Archaea within two-phase biogas reactor systems operated with plant biomass.

The two-phase leach-bed system is a biogas reactor system optimized for the utilization of energy crop silages at maximized loading rates under maintenance of an optimal microbial activity. In this study, a characterization of the methanogenic microbial community within this reactor system was conducted for the first time. Accordingly, effluent samples from the anaerobic filter and the silage digesting leach-bed reactors of both a laboratory-scale two-phase biogas reactor system and a scaled-up commercial on-farm pilot plant were investigated. In total, five Archaea-specific 16S rDNA libraries were constructed and analyzed by amplified rDNA restriction analysis (ARDRA), with subsequent phylogenetic analysis of nucleotide sequences for individual ARDRA patterns. A quantification of major methanogenic Archaea groups was conducted by real-time PCR. A total of 663 clones were analyzed and 45 operational taxonomic units (OTUs) related to methanogenic Archaea were detected. These OTUs were related to the orders Methanosarcinales, Methanomicrobiales and Methanobacteriales, as well as the hitherto uncultured CA-11 and ARC-I groups, and most of them occurred throughout all the compartments of both two-phase biogas reactors. The proportion of acetotrophic to hydrogenotrophic methanogens differed between the laboratory and the pilot scale system. A total of 56% of the clones from the 16S rDNA library derived from the laboratory biogas system were assigned to presumably acetotrophic members of Methanosarcinales. In contrast, these OTUs were less abundant in the 16S rDNA library derived from samples of the pilot plant. Therein, the most dominant OTUs were Methanoculleus-related OTUs, which presumably indicated the predominant presence of hydrogenotrophic methanogens. These findings were confirmed by group-specific quantitative real-time PCR assays. The results indicated that the fraction of acetotrophic and hydrogenotrophic methanogens within a biogas reactor caused certain variations, which may reflect varying substrate utilization during methanogenesis.

Toxicity screening of biochar-mineral composites using germination tests.

This study assessed the properties and toxicity (water cress germination trials) of 38 waste-derived, novel biochar-mineral composites (BMCs) produced via slow pyrolysis and hydrothermal carbonization (hydrochars). The biochars were produced from sewage sludge and compost-like output (CLO) by varying the type of mineral additive (zeolite, wood ash and lignite fly ash), the mineral-to-feedstock ratio and the carbonization process. While pure hydrochars completely inhibited germination of water cress, this effect was ameliorated by mineral additives. Seedlings grew best in pyrolysis chars and while wood ash addition decreased plant growth in many cases, 1:10 addition to CLO doubled germination rate. The factors responsible for the phytotoxicity can be attributed to pH, salinity and organic contaminants. Importantly, while pure minerals inhibited germination, conversion of minerals into BMCs reduced their inhibitory effects due to buffered release of minerals. Overall, mineral wastes (e.g., combustion ashes) and waste biomass can be used safely as sources of nutrients and stable organic carbon (for soil carbon sequestration) when converted into specific biochar-mineral composites, exploiting synergies between the constituents to deliver superior performance.

A biologically inspired variable-pH strategy for enhancing short-chain fatty acids (SCFAs) accumulation in maize straw fermentation.

This study investigates the feasibility of varying the pH to enhance the accumulation of short-chain fatty acids (SCFAs) in the in vitro fermentation of maize straw. The corresponding hydrolysis rate and the net SCFA yield increased as inoculum ratio (VSinoculum/VSsubstrate) increased from 0.09 to 0.79. The pH were maintained at 5.3, 5.8, 6.3, 6.8, 7.3, and 7.8, respectively. A neutral pH of approximately 6.8 was optimal for hydrolysis. The net SCFA yield decreased by 34.9% for a pH of less than 5.8, but remained constant at approximately 721±5mg/gvs for a pH between 5.8 and 7.8. In addition, results were obtained for variable and constant pH levels at initial substrate concentrations of 10, 30 and 50g/L. A variable pH increased the net SCFA yield by 23.6%, 29.0%, and 36.6% for concentrations of 10, 30 and 50g/L. Therefore, a variable pH enhanced SCFA accumulation in maize straw fermentation.

Toward the Standardization of Biochar Analysis: The COST Action TD1107 Interlaboratory Comparison.

Biochar produced by pyrolysis of organic residues is increasingly used for soil amendment and many other applications. However, analytical methods for its physical and chemical characterization are yet far from being specifically adapted, optimized, and standardized. Therefore, COST Action TD1107 conducted an interlaboratory comparison in which 22 laboratories from 12 countries analyzed three different types of biochar for 38 physical-chemical parameters (macro- and microelements, heavy metals, polycyclic aromatic hydrocarbons, pH, electrical conductivity, and specific surface area) with their preferential methods. The data were evaluated in detail using professional interlaboratory testing software. Whereas intralaboratory repeatability was generally good or at least acceptable, interlaboratory reproducibility was mostly not (20% < mean reproducibility standard deviation < 460%). This paper contributes to better comparability of biochar data published already and provides recommendations to improve and harmonize specific methods for biochar analysis in the future.

Integration of pyrolysis and anaerobic digestion--use of aqueous liquor from digestate pyrolysis for biogas production.

Anaerobic digestion of aqueous pyrolysis liquor derived from pyrolysis of solid digestate was tested in batch mode using an un-adapted inoculum. Three pyrolysis liquors produced at 330°C, 430°C and 530°C in four COD-based concentrations of 3, 6, 12 and 30 g L(-1) were investigated. The three lower concentrations showed considerable biogas production, whereas the 30 g L(-1) dosage caused process inhibition. The highest methane yield of 199.1±18.5 mL g(COD)(-1) (COD removal: 56.9±5.3%) was observed for the 330°C pyrolysis liquor, followed by the 430°C sample with only slightly lower values. The 530°C sample dropped to a yield of 129.3±19.7 mL g(COD)(-1) (COD removal: 36.9±5.6%). Most VOCs contained in the pyrolysis liquor (i.e. furfural, phenol, catechol, guaiacol, and levoglucosan) were reduced below detection limit (cresol by 10-60%). Consequently, integrated pyrolysis and anaerobic digestion in addition to thermochemical conversion of digestate also promises bioconversion of pyrolysis liquors.

Influence of digestion temperature and organic loading rate on the continuous anaerobic treatment of process liquor from hydrothermal carbonization of sewage sludge.

This experimental study investigates the use of process liquor from hydrothermal carbonization (HTC) of sewage sludge as sole substrate for anaerobic digestion (AD). The process performance at 37°C (mesophilic) and 55°C (thermophilic) was determined based on two identical continuously-fed anaerobic filters (26 L each) operated for 20 weeks. During operation, the organic loading rate was stepwise increased from 1 to 5 gCOD L(-1)d(-)(1). This decreased the hydraulic retention time from 34 to 5 days. Significant differences in methane production were not observed as both reactors yielded up to 0.18 LCH4 gCOD(-)(1). Increased temperature had no effect on the steady-state COD removal efficiency with both reactors stabilized at 68-75%. Macro- and micronutrients were provided by HTC liquor in sufficient concentrations throughout the experiment. Methanogenesis was identified as the speed-limiting step in anaerobic digestion of HTC liquor. Hydrolysis was of an order of magnitude faster than methanogenesis.

Upflow anaerobic solid-state (UASS) digestion of horse manure: Thermophilic vs. mesophilic performance.

Energetic use of complex lignocellulosic wastes has gained global interest. Thermophilic digestion of horse manure based on straw was investigated using the upflow anaerobic solid-state (UASS) process. Increasing the organic loading rate from 2.5 to 5.5gvsL(-)(1)d(-)(1) enhanced the average methane production rate from 0.387 to 0.687LCH4L(-)(1)d(-)(1), whereas the yield decreased from 154.8 to 124.8LCH4kgvs(-)(1). A single-stage and two-stage process design showed almost the same performance. Compared to prior experiments at mesophilic conditions, thermophilic conditions showed a significantly higher efficiency with an increase of 59.8% in methane yield and 58.1% in methane production rate. Additional biochemical methane potential (BMP) tests with two types of horse manure and four different bedding materials showed that wheat straw obtained the highest BMP. The results show that the thermophilic UASS process can be the key to an efficient energy recovery from straw-based manures.

Characterization of products from hydrothermal carbonization of orange pomace including anaerobic digestibility of process liquor.

In this study, the effect of the temperature and reaction time on hydrothermal carbonization (HTC) of orange pomace was investigated. In addition, a set of anaerobic batch tests were performed to determine the resulting biogas and methane potential of the spent liquor. Hydrochar yields followed a decreasing trend with the increasing temperature, whereas reaction time had no considerably effect on the yield. The evolution of the H/C and O/C atomic ratios from the raw material to the hydrochars suggested that dehydration reactions prevail during HTC. The hydrochars tended to become enriched in Ca, Mg and P minerals by increasing HTC temperature. The heavy metal contents in hydrochars were found below limits and no PAH compound was detected. Anaerobic digestion tests showed that the aqueous phase from HTC can be used as feedstocks for biogas production.

Production, characterization, and biogas application of magnetic hydrochar from cellulose.

Hydrothermal carbonization (HTC) produces carbon-rich nano-micro size particles. In this study, magnetic hydrochar (MHC) was prepared from model compound cellulose by simply adding ferrites during HTC. The effects of ferrites on HTC were evaluated by characterizing solid MHC and corresponding process liquid. Additionally, magnetic stability of MHC was tested by magnetic susceptibility method. Finally, MHC was used as support media for anaerobic films in anaerobic digestion (AD). Ash-free mass yield was around 50% less in MHC than hydrochar produced without ferrites at any certain HTC reaction condition, where organic part of MHC is mainly carbon. In fact, amorphous hydrochar was growing on the surface of inorganic ferrites. MHC maintained magnetic susceptibility regardless of reaction time at reaction temperature 250°C. Pronounced inhibitory effects of magnetic hydrochar occurred during start-up of AD but diminished with prolong AD times. Visible biofilms were observed on the MHC by laser scanning microscope after AD.

Harmonization of the quantitative determination of volatile fatty acids profile in aqueous matrix samples by direct injection using gas chromatography and high-performance liquid chromatography techniques: Multi-laboratory validation study.

The performance parameters of volatile fatty acids (VFAs) measurements were assessed for the first time by a multi-laboratory validation study among 13 laboratories. Two chromatographic techniques (GC and HPLC) and two quantification methods such as external and internal standard (ESTD/ISTD) were combined in three different methodologies GC/ESTD, HPLC/ESTD and GC/ISTD. Linearity evaluation of the calibration functions in a wide concentration range (10-1000mg/L) was carried out using different statistical parameters for the goodness of fit. Both chromatographic techniques were considered similarly accurate. The use of GC/ISTD, despite showing similar analytical performance to the other methodologies, can be considered useful for the harmonization of VFAs analytical methodology taking into account the normalization of slope values used for the calculation of VFAs concentrations. Acceptance criteria for VFAs performance parameters of the multi-laboratory validation study should be established as follows: (1) instrument precision (RSDINST≤1.5%); (2) linearity (R(2)≥0.998; RSDSENSITIVITY≤4%; REMAX≤8%; REAVER≤ 3%); (3) precision (RSD≤1.5%); (4) trueness (recovery of 97-103%); (5) LOD (≤3mg/L); and (6) LOQ (10mg/L).

Use of biochars in anaerobic digestion.

This study investigated the behavior of biochars from pyrolysis (pyrochar) and hydrothermal carbonization (hydrochar) in anaerobic digestion regarding their degradability and their effects on biogas production and ammonia inhibition. A batch fermentation experiment (42°C, 63 days) was conducted in 100mL syringes filled with 30 g inoculum, 2g biochar and four levels of total ammonium nitrogen (TAN). For pyrochar, no clear effect on biogas production was observed, whereas hydrochar increased the methane yield by 32%. This correlates with the hydrochar's larger fraction of anaerobically degradable carbon (10.4% of total carbon, pyrochar: 0.6%). Kinetic and microbiota analyses revealed that pyrochar can prevent mild ammonia inhibition (2.1 g TANk g(-1)). Stronger inhibitions (3.1-6.6 g TAN kg(-1)) were not mitigated, neither by pyrochar nor by hydrochar. Future research should pay attention to biochar-microbe interactions and the effects in continuously-fed anaerobic digesters.

Evaluation of integrated anaerobic digestion and hydrothermal carbonization for bioenergy production.

Lignocellulosic biomass is one of the most abundant yet underutilized renewable energy resources. Both anaerobic digestion (AD) and hydrothermal carbonization (HTC) are promising technologies for bioenergy production from biomass in terms of biogas and HTC biochar, respectively. In this study, the combination of AD and HTC is proposed to increase overall bioenergy production. Wheat straw was anaerobically digested in a novel upflow anaerobic solid state reactor (UASS) in both mesophilic (37 °C) and thermophilic (55 °C) conditions. Wet digested from thermophilic AD was hydrothermally carbonized at 230 °C for 6 hr for HTC biochar production. At thermophilic temperature, the UASS system yields an average of 165 LCH4/kgVS (VS: volatile solids) and 121 L CH4/kgVS at mesophilic AD over the continuous operation of 200 days. Meanwhile, 43.4 g of HTC biochar with 29.6 MJ/kgdry_biochar was obtained from HTC of 1 kg digestate (dry basis) from mesophilic AD. The combination of AD and HTC, in this particular set of experiment yield 13.2 MJ of energy per 1 kg of dry wheat straw, which is at least 20% higher than HTC alone and 60.2% higher than AD only.

Hydrothermal carbonization (HTC): near infrared spectroscopy and partial least-squares regression for determination of selective components in HTC solid and liquid products derived from maize silage.

Near-infrared (NIR) spectroscopy was evaluated as a rapid method of predicting fiber components (hemicellulose, cellulose, lignin, and ash) and selective compounds of hydrochar and corresponding process liquor produced by hydrothermal carbonization (HTC) of maize silage. Several HTC reaction times and temperatures were applied and NIR spectra of both HTC solids and liquids were obtained and correlated with concentration determined from van-Soest fiber analysis, IC, and UHPLC. Partial least-squares regression was applied to calculate models for the prediction of selective substances. The model developed with the spectra had the best performance in 3-7 factors with a correlation coefficient, which varied between 0.9275-0.9880 and 0.9364-0.9957 for compounds in solid and liquid, respectively. Calculated root mean square errors of prediction (RMSEP) were 0.42-5.06mg/kg. The preliminary results indicate that NIR, a widely applied technique, might be applied to determine chemical compounds in HTC solid and liquid.

Anaerobic digestion of horse dung mixed with different bedding materials in an upflow solid-state (UASS) reactor at mesophilic conditions.

Aim of this study was to investigate the use of upflow anaerobic solid-state (UASS) digestion for treating horse manure. Biochemical methane potential (BMP) tests conducted for varying mixtures of dung (hay and silage feed) and bedding material (wheat straw, flax, hemp, wood chips) showed that straw mixed with hay horse dung has the highest potential of [Formula: see text] . Continuous mesophilic digestion was conducted for 238 days using a single-stage UASS reactor (27 L) and a two-stage UASS system with an anaerobic filter (AF, 21 L). Increasing the organic loading rate (OLR) from 2.5 to 4.5 g vs L(-1)d(-1) enhanced the methane rate of the single-stage reactor from 0.262 to 0.391 LL(-1)d(-1) while the methane yield declined from 104.8 to 86.9 L kg vs(-1). The two-stage system showed similar yields. Thus, for solid-state digestion of horse manure a single-stage UASS reactor appears sufficient.

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.

Anaerobic digestion of wheat straw--performance of continuous solid-state digestion.

In this study the upflow anaerobic solid-state (UASS) reactor was operated at various conditions to optimize the process parameters for anaerobically digesting wheat straw in a continuous process. Additionally, particle size effects have been studied in the operation at 55 and 60°C. Moreover, the incremental effect of the organic loading rate (OLR) to the system was examined from 2.5 to 8 gVS L(-1) d(-1). It was found that the UASS operating at 60 °C with a small OLR yields highest methane production, but the advantage over thermophilic operation is negligible. The rise in OLR reduces the systems yields, as expected. From OLR=8 gVS L(-1) d(-1) a second stage is necessary to circumvent volatile fatty acids accumulation.

Chemical and morphological changes in hydrochars derived from microcrystalline cellulose and investigated by chromatographic, spectroscopic and adsorption techniques.

Hydrothermal carbonization (HTC) can be used for converting the biomass into a carbon-rich material, whose application as a fuel requires higher heating value, whereas soil amendment needs stable carbon. This work was focused on the characterization of hydrochars derived from microcrystalline cellulose. The chars were investigated using elemental analysis, Brunauer-Emmett-Teller technique, nuclear magnetic resonance spectroscopy, Raman, Fourier transform infrared, and electron spin resonance spectroscopy. Severity in temperature between 230 and 270°C with reaction times between 2 and 10 h only affect the carbon content moderately. The results show that aromatization of HTC chars correlates well with temperature, which was further supported by the increase of organic radicals with decreasing g values at higher temperatures. Based on these results, the energetic use of chars favors mild HTC (T<230°C and t≤6 h), while the soil amendement favors serve conditions (T≥230°C, and t>6 h).