Viability of Wildflower Seeds After Mesophilic Anaerobic Digestion in Lab-Scale Biogas Reactors.

The use of wildflower species as biogas feedstock carries the risk that their seeds survive anaerobic digestion (AD) and cause weed problems if spread with the digestate. Risk factors for seed survival in AD include low temperature, short exposure and hardseededness (HS). However, it is not possible to predict how AD will affect seed viability of previously unstudied species. In laboratory-scale reactors, we exposed seeds of eight species from a mixture of flowering wild plants intended as biogas feedstock and three reference species to AD at two mesophilic temperatures. Half of the species were HS, the other was non-HS (NHS). Viability was determined using a combination of tetrazolium and germination tests. Viability and germinability were modeled as functions of exposure time using a dose-response approach. Responses to AD varied considerably among species, and none of the considered influencing factors (time, temperature, HS) had a consistent effect. Seed lots of a species differed in inactivation times and seed-killing efficacy. The HS species Melilotus officinalis, Melilotus albus, and Malva sylvestris were particularly AD-resistant. They were the only ones that exhibited biphasic viability curves and tended to survive and germinate more at 42°C than at 35°C. Viability of the remaining species declined in a sigmoidal curve. Most NHS species were inactivated within a few days (Cichorium intybus, Daucus carota, Echium vulgare, and Verbascum thapsus), while HS species survived longer (Malva alcea). AD stimulated germination in the HS species A. theophrasti and its AD-resistance overlapped with that of the most resistant NHS species, C. album and tomato. In all seed lots, germinability was lost faster than viability, implying that mainly dormant seeds survived. After the maximum exposure time of 36 days, seeds of HS species and Chenopodium album were still viable. We concluded that viability responses to mesophilic AD were determined by the interplay of AD-conditions and species- and seed-lot-specific traits, of which HS was an important but only one factor. For the use of wildflowers as biogas feedstock, we recommended long retention times and special care with regard to HS species.

Chemical and Enzymatic Synthesis of Biobased Xylo-Oligosaccharides and Fermentable Sugars from Wheat Straw for Food Applications.

Xylo-oligosaccharides are sugar oligomers with 2~7 xylose units considered non-digestible fibers that can be produced from biodegradable and low-cost biomass like wheat straw. An integrated approach consisting of hydrothermal pretreatment, alkaline treatment, enzymatic treatment and the combinations thereof was applied to overcome the recalcitrance structure of the wheat straw and allow selective fractioning into fermentable sugars and xylo-oligosaccharides. The hydrolysates and processed solids were chemically characterized by High-performance liquid chromatography and Ion chromatography, and the results were expressed as function of the severity factor and statistically interpreted. The concentration of fermentable sugars (glucose, xylose, arabinose) was the highest after the combination of alkaline and enzymatic treatment with xylanase (18 g/L sugars), while xylo-oligosaccharides (xylotriose and xylotetraose) were released in lower amounts (1.33 g/L) after the same treatment. Refining experiments were carried out to obtain a purified fraction by using anion and cation exchange chromatography. The polymer adsorber resin MN-502 showed efficient removal of salts, phenols and furan derivatives. However, the xylo-oligosaccharides yields were also slightly reduced. Although still requiring further optimization of the treatments to obtain higher purified oligomer yields, the results provide information on the production of xylo-oligosaccharides and fermentable sugars from wheat straw for potential use in food applications.

Marker microbiome clusters are determined by operational parameters and specific key taxa combinations in anaerobic digestion.

In this study, microbiomes of 36 full-scale anaerobic digesters originated from 22 different biogas plants were compared by terminal restriction fragment length polymorphism (TRFLP) analysis. Regarding the differences in microbial community composition, a weighting of the environmental parameters could be derived from higher to lower importance as follows: (i) temperature, (ii) TAN and NH3 concentrations and conductivity, and (iii) the chemical composition of the supplied feedstocks. Biotic interactions between specific bacterial and archaeal community arrangements were revealed, whereby members of the phyla Bacteroidetes and Cloacimonetes combined with the archaeal genus Methanothrix dominated the conversion of homogeneous feedstocks, such as waste water sludge or industrial waste. As most of the detected TRFs were only found in a certain number of anaerobic digestion plants, each plant develops its unique microbiome. The putative rare species, the specialists, are potentially hidden drivers of microbiome functioning as they provide necessary traits under, e.g., process-inconvenient conditions.

Biogas crops grown in energy crop rotations: Linking chemical composition and methane production characteristics.

Methane production characteristics and chemical composition of 405 silages from 43 different crop species were examined using uniform laboratory methods, with the aim to characterise a wide range of crop feedstocks from energy crop rotations and to identify main parameters that influence biomass quality for biogas production. Methane formation was analysed from chopped and over 90 days ensiled crop biomass in batch anaerobic digestion tests without further pre-treatment. Lignin content of crop biomass was found to be the most significant explanatory variable for specific methane yields while the methane content and methane production rates were mainly affected by the content of nitrogen-free extracts and neutral detergent fibre, respectively. The accumulation of butyric acid and alcohols during the ensiling process had significant impact on specific methane yields and methane contents of crop silages. It is proposed that products of silage fermentation should be considered when evaluating crop silages for biogas production.

Energy balance, greenhouse gas emissions, and profitability of thermobarical pretreatment of cattle waste in anaerobic digestion.

In this study modeled full scale application of thermobarical hydrolysis of less degradable feedstock for biomethanation was assessed in terms of energy balance, greenhouse gas emissions, and economy. Data were provided whether the substitution of maize silage as feedstock for biogas production by pretreated cattle wastes is beneficial in full-scale application or not. A model device for thermobarical treatment has been suggested for and theoretically integrated in a biogas plant. The assessment considered the replacement of maize silage as feedstock with liquid and/or solid cattle waste (feces, litter, and feed residues from animal husbandry of high-performance dairy cattle, dry cows, and heifers). The integration of thermobarical pretreatment is beneficial for raw material with high contents of organic dry matter and ligno-cellulose: Solid cattle waste revealed very short payback times, e.g. 9 months for energy, 3 months for greenhouse gases, and 3 years 3 months for economic amortization, whereas, in contrast, liquid cattle waste did not perform positive replacement effects in this analysis.

Hygiene and Sanitation in Biogas Plants.

The increasing number of agricultural biogas plants and higher amounts of digestate spread on agricultural land arouse a considerable interest in the hygiene situation of digested products. This chapter reviews the current knowledge on sanitation during anaerobic digestion and the hygienic status of digestate concerning a multitude of pathogens potentially compromising the health of humans, animals and plants. Physical, chemical and biological parameters influencing the efficiency of sanitation in anaerobic digestion are considered. The degree of germ reduction depends particularly on the resistance of the pathogen of concern, the processing conditions, the feedstock composition and the diligence of the operation management. Most scientific studies facing sanitation in biogas plants have provided data ascertaining reduction of pathogens by the biogas process. Some pathogens, however, are able to persist virtually unaffected due to the ability to build resistant permanent forms. As compared to the feedstock, the sanitary status of the digestate is thus improved or in the worst case, the sanitary quality remains almost unchanged. According to this, the spreading of digestate on agricultural area in accordance to current rules and best practice recommendations is considered to impose no additional risk for the health of humans, animals and plants.

Improving aerobic stability and biogas production of maize silage using silage additives.

The effects of air stress during storage, exposure to air at feed-out, and treatment with silage additives to enhance aerobic stability on methane production from maize silage were investigated at laboratory scale. Up to 17% of the methane potential of maize without additive was lost during seven days exposure to air on feed-out. Air stress during storage reduced aerobic stability and further increased methane losses. A chemical additive containing salts of benzoate and propionate, and inoculants containing heterofermentative lactic acid bacteria were effective to increase aerobic stability and resulted in up to 29% higher methane yields after exposure to air. Exclusion of air to the best possible extent and high aerobic stabilities should be primary objectives when ensiling biogas feedstocks.

Community shifts in a well-operating agricultural biogas plant: how process variations are handled by the microbiome.

This study provides a comprehensive, long-term microbiological study of a continuously operated, mesophilic, agricultural biogas plant fed with whole-crop silages of maize and rye, cattle manure and cattle slurry. The microbial community structure was accessed by high-throughput 16S rRNA gene amplicon sequencing. For the characterisation of the microbial dynamics, the community profiling method terminal restriction fragment length polymorphism (TRFLP) in combination with a cloning-sequencing approach as well as a LC-MS/MS approach for protein identification were applied. Our results revealed that the anaerobic digestion is a highly sensitive process: small variations in the process performance induce fluctuations in the microbial community composition and activity. In this context, it could be proven that certain microbial species were better adapted to changing process condition such as temperature (interspecies competition) and that there is a physiological compensation between different microorganisms so that the reactor efficiency was not adversely affected despite of structural and functional changes within the microbial community.

Effect of liquid hot water pre-treatment on sugarcane press mud methane yield.

Sugarcane press mud was pretreated by liquid hot water (LHW) at different temperatures (140-210 °C) and pre-treatment times (5-20 min) in order to assess the effects on the chemical oxygen demand (COD) solubilisation, inhibitors formation and methane yield. The experimental results showed that a high degree of biomass solubilisation was possible using LHW. Higher methane yields were obtained at lower severities (log(Ro) = 2.17-2.77) with (i) mild temperatures (140-150 °C) and long contact times (12.5 min, 20 min) or (ii) mild temperatures (175 °C) with short contact time (2 min). The highest increase in methane yield (up to 63%) compared to the untreated press mud was found at 150 °C for 20 min. At temperatures of 200 °C and 210 °C, low methane efficiency was attributed to the possible formation of refractory compounds through the Maillard reaction.

Effects of thermobarical pretreatment of cattle waste as feedstock for anaerobic digestion.

Lab-scale experiments were conducted to assess the impact of thermobarical treatment of cattle waste on anaerobic digestion. Treatment was at temperatures of 140-220°C in 20K steps for a 5-min duration. Methane yields could be increased by up to 58% at a treatment temperature of 180°C. At 220°C the abundance of inhibitors and other non-digestible substances led to lower methane yields than those obtained from untreated material. In an extended analysis it could be demonstrated that there is a functional correlation between the methane yields after 30 days and the formation rate and methane yield in the acceleration phase. It could be proved in a regression of these correlation values that the optimum treatment temperature is 164°C and that the minimum treatment temperature should be above 115°C.

Kinetics of inactivation and dilution effects on the mass balance of fungal phytopathogens in anaerobic digesters.

Knowledge of fate and behavior of plant pathogens in the biogas production chain is limited and hampers the estimation and evaluation of the potential phytosanitary risk if digestate is spread on arable land as a fertilizer. Therefore, simulation is an appropriate tool to demonstrate the effects which influence the steady state of pathogen infected plant material in both digesters and digestate. Simple approaches of kinetics of inactivation and mass balances of infected material were carried out considering single-step as well as two-step digestion. The simulation revealed a very fast to fast reduction of infected material after a singular feeding, reaching a cutback to less than 1% of input within 4 days even for D90-values of 68 h. Steady state mass balances below input rate could be calculated with D90-values of less than 2 h at a continuous hourly feeding. At higher D90-values steady state mass balances exceed the input rate but are still clearly below the sum of input mass. Dilution further decreases mass balances to values 10(-5) to 10(-6) Mg m(-3) for first-step digestion and 10(-8) to 10(-9) for second-step.

Effects of ensiling, silage additives and storage period on methane formation of biogas crops.

Effects of the ensiling process, storage periods of up to 1 year and several chemical and biological silage additives on biomethanation were assessed for maize, sorghum, forage rye and triticale with the aim to identify optimised conditions for silage production of crops used as feedstock in biogas plants. Ensiling, prolonged storage and biological silage additives showed positive effects on methane yield of up to 11%. These could be attributed to increases in organic acids and alcohols during ensiling. A regression model including acetic acid, butyric acid and ethanol accounts for 75-96% of the variation in methane yield. Storage periods of up to 1 year for properly ensiled crops could be possible without losses in methane production, considering the increase in methane yield and the losses of dry matter during this period. However, taking storage losses into account silage additives showed little effect on methane production.

Concepts and profitability of biogas production from landscape management grass.

Landscape management grass is generally harvested late, resulting in unfavorable composition for many utilization purposes. This study explores various technical concepts of biogas production and their economic viability. The Lower Oder Valley National Park is taken here as an example. This National Park in North-East Germany comprises large grassland areas with conservation-related restrictions on management. The concepts of biogas production and use considered are: (1) decentralized digestion and use of biogas at five autonomous combined heat and power (CHP) units, (2) decentralized digestion and delivery of the biogas to a centralized CHP unit, (3) decentralized digestion, upgrading of the biogas and feeding into the natural gas grid, and (4) one central biogas plant with centralized CHP unit. Annual costs and revenues of biogas production were calculated for each alternative. Biogas production from landscape management grass meets the conservational demands of late cutting periods and under certain circumstances shows a profit.