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    DNA and RNA extraction and quantitative real-time PCR-based assays for biogas biocenoses in an interlaboratory comparison
    (Basel : MDPI, 2016) Lebuhn, Michael; Derenkó, Jaqueline; Rademacher, Antje; Helbig, Susanne; Munk, Bernhard; Pechtl, Alexander; Stolze, Yvonne; Prowe, Steffen; Schwarz, Wolfgang H.; Schlüter, Andreas; Liebl, Wolfgang; Klocke, Michael
    Five institutional partners participated in an interlaboratory comparison of nucleic acid extraction, RNA preservation and quantitative Real-Time PCR (qPCR) based assays for biogas biocenoses derived from different grass silage digesting laboratory and pilot scale fermenters. A kit format DNA extraction system based on physical and chemical lysis with excellent extraction efficiency yielded highly reproducible results among the partners and clearly outperformed a traditional CTAB/chloroform/isoamylalcohol based method. Analytical purpose, sample texture, consistency and upstream pretreatment steps determine the modifications that should be applied to achieve maximum efficiency in the trade-off between extract purity and nucleic acid recovery rate. RNA extraction was much more variable, and the destination of the extract determines the method to be used. RNA stabilization with quaternary ammonium salts was an as satisfactory approach as flash freezing in liquid N2. Due to co-eluted impurities, spectrophotometry proved to be of limited value for nucleic acid qualification and quantification in extracts obtained with the kit, and picoGreen® based quantification was more trustworthy. Absorbance at 230 nm can be extremely high in the presence of certain chaotropic guanidine salts, but guanidinium isothiocyanate does not affect (q)PCR. Absolute quantification by qPCR requires application of a reliable internal standard for which correct PCR efficiency and Y-intercept values are important and must be reported.
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    Uncertainty in the measurement of indoor temperature and humidity in naturally ventilated dairy buildings as influenced by measurement technique and data variability
    (Amsterdam : Elsevier, 2017) Hempel, Sabrina; König, Marcel; Menz, Christoph; Janke, David; Amon, Barbara; Banhazi, Thomas M.; Estellés, Fernando; Amon, Thomas
    The microclimatic conditions in dairy buildings affect animal welfare and gaseous emissions. Measurements are highly variable due to the inhomogeneous distribution of heat and humidity sources (related to farm management) and the turbulent inflow (associated with meteorologic boundary conditions). The selection of the measurement strategy (number and position of the sensors) and the analysis methodology adds to the uncertainty of the applied measurement technique. To assess the suitability of different sensor positions, in situations where monitoring in the direct vicinity of the animals is not possible, we collected long-term data in two naturally ventilated dairy barns in Germany between March 2015 and April 2016 (horizontal and vertical profiles with 10 to 5 min temporal resolution). Uncertainties related to the measurement setup were assessed by comparing the device outputs under lab conditions after the on-farm experiments. We found out that the uncertainty in measurements of relative humidity is of particular importance when assessing heat stress risk and resulting economic losses in terms of temperature-humidity index. Measurements at a height of approximately 3 m–3.5 m turned out to be a good approximation for the microclimatic conditions in the animal occupied zone (including the air volume close to the emission active zone). However, further investigation along this cross-section is required to reduce uncertainties related to the inhomogeneous distribution of humidity. In addition, a regular sound cleaning (and if possible recalibration after few months) of the measurement devices is crucial to reduce the instrumentation uncertainty in long-term monitoring of relative humidity in dairy barns. © 2017 The Authors
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    Kaskadennutzung von Lignocellulose : LX-Verfahren trifft auf B. coagulans
    (Heidelberg : Spektrum, 2020) Schroedter, Linda; Streffer, Friedrich; Streffer, Katrin; Unger, Peter; Venus, Joachim
    Investigating alternatives for petrobased substrates, lignocellulose is an interesting yet complex feedstock that offers various possibilities for the design of new and sustainable chemical routes. The novel energy-saving LX-pretreatment was combined with thermophilic Bacillus coagulans. By this, corn straw was used in an innovative cascade obtaining biogas, lignin as well as polymerisable L-(+)-lactic acid of over 99 percents optical purity. © 2020, Die Autoren.
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    Influence of Tree Species, Harvesting Method and Storage on Energy Demand and Wood Chip Quality When Chipping Poplar, Willow and Black Locust
    (Basel : MDPI AG, 2020) Pecenka, Ralf; Lenz, Hannes; Jekayinfa, Simeon Olatayo; Hoffmann, Thomas
    The cultivation of fast-growing wood (e.g., poplar, willow or black locust) in short rotation coppices and agroforestry systems presents an opportunity for producing biomass sustainably in the agricultural sector. Cost-efficient agricultural wood production requires the availability of high-performance machinery and methods with which high-quality wood chips can be produced at low cost. It is known from harvesting short rotation coppices in practice that both the wood chip quality and the performance of the harvesting machinery depend on a variety of factors (e.g., harvesting method, weather conditions, tree species). That is why this study examines in detail the influence of the tree species (different varieties of poplar, willow, black locust) and the wood condition (fresh, stored or dried, frozen) on the specific energy demand for comminution in a stationary drum chipper and on the particle size distribution of the wood chips produced. For all the tree species examined, the chipping of dried as well as frozen stems was connected with a significant increase in the specific energy demand for comminution. An increase of 31% has been measured if poplar stems are chipped in frozen conditions (max. 6.31 kWh t−1). Drying led to an increase of 59% for dried willow stems (max. 6.67 kWh t−1). Drying and frost had also an influence on the size and quality of the wood chips, but no globally significant connection could be established for the examined tree varieties.
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    Effect of Fans’ Placement on the Indoor Thermal Environment of Typical Tunnel-Ventilated Multi-Floor Pig Buildings Using Numerical Simulation
    (Basel : MDPI AG, 2022) Wang, Xiaoshuai; Cao, Mengbing; Hu, Feiyue; Yi, Qianying; Amon, Thomas; Janke, David; Xie, Tian; Zhang, Guoqiang; Wang, Kaiying
    An increasing number of large pig farms are being built in multi-floor pig buildings (MFPBs) in China. Currently, the ventilation system of MFPB varies greatly and lacks common standards. This work aims to compare the ventilation performance of three popular MFPB types with different placement of fans using the Computational Fluid Dynamics (CFD) technique. After being validated with field-measured data, the CFD models were extended to simulate the air velocity, air temperature, humidity, and effective temperature of the three MFPBs. The simulation results showed that the ventilation rate of the building with outflowing openings in the endwall and fans installed on the top of the shaft was approximately 25% less than the two buildings with fans installed on each floor. The ventilation rate of each floor increased from the first to the top floor for both buildings with a shaft, while no significant difference was observed in the building without a shaft. Increasing the shaft’s width could mitigate the variation in the ventilation rate of each floor. The effective temperature distribution at the animal level was consistent with the air velocity distribution. Therefore, in terms of the indoor environmental condition, the fans were recommended to be installed separately on each floor.
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    Impact of Camera Viewing Angle for Estimating Leaf Parameters of Wheat Plants from 3D Point Clouds
    (Basel : MDPI, 2021) Li, Minhui; Shamshiri, Redmond R.; Schirrmann, Michael; Weltzien, Cornelia
    Estimation of plant canopy using low-altitude imagery can help monitor the normal growth status of crops and is highly beneficial for various digital farming applications such as precision crop protection. However, extracting 3D canopy information from raw images requires studying the effect of sensor viewing angle by taking into accounts the limitations of the mobile platform routes inside the field. The main objective of this research was to estimate wheat (Triticum aestivum L.) leaf parameters, including leaf length and width, from the 3D model representation of the plants. For this purpose, experiments with different camera viewing angles were conducted to find the optimum setup of a mono-camera system that would result in the best 3D point clouds. The angle-control analytical study was conducted on a four-row wheat plot with a row spacing of 0.17 m and with two seeding densities and growth stages as factors. Nadir and six oblique view image datasets were acquired from the plot with 88% overlapping and were then reconstructed to point clouds using Structure from Motion (SfM) and Multi-View Stereo (MVS) methods. Point clouds were first categorized into three classes as wheat canopy, soil background, and experimental plot. The wheat canopy class was then used to extract leaf parameters, which were then compared with those values from manual measurements. The comparison between results showed that (i) multiple-view dataset provided the best estimation for leaf length and leaf width, (ii) among the single-view dataset, canopy, and leaf parameters were best modeled with angles vertically at -45⸰_ and horizontally at 0⸰_ (VA -45, HA 0), while (iii) in nadir view, fewer underlying 3D points were obtained with a missing leaf rate of 70%. It was concluded that oblique imagery is a promising approach to effectively estimate wheat canopy 3D representation with SfM-MVS using a single camera platform for crop monitoring. This study contributes to the improvement of the proximal sensing platform for crop health assessment. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
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    Using SPOT-7 for Nitrogen Fertilizer Management in Oil Palm
    (Basel : MDPI AG, 2020) Yadegari, Mohammad; Shamshiri, Redmond R.; Shariff, Abdul Rashid Mohamed; Balasundram, Siva K.; Mahns, Benjamin
    Environmental concerns are growing about excessive applying nitrogen (N) fertilizers, especially in oil palm. Some conventional methods which are used to assess the amount of nutrient in oil palm are time-consuming, expensive, and involve frond destruction. Remote sensing as a non-destructive, affordable, and efficient method is widely used to detect the concentration of chlorophyll (Chl) from canopy plants using several vegetation indices (VIs) because there is an influential relation between the concentration of N in the leaves and canopy Chl content. The objectives of this research are to (i) evaluate and compare the performance of various vegetation indices (VIs) for measuring N status in oil palm canopy using SPOT-7 imagery (AIRBUS Defence & Space, Ottobrunn, Germany) to (ii) develop a regression formula that can predict the N content using satellite data to (iii) assess the regression formula performance on testing datasets by testing the coefficient of determination between the predicted and measured N contents. SPOT-7 was acquired in a 6-ha oil palm planted area in Pahang, Malaysia. To predict N content, 28 VIs based on the spectral range of SPOT-7 satellite images were evaluated. Several regression models were applied to determine the highest coefficient of determination between VIs and actual N content from leaf sampling. The modified soil-adjusted vegetation index (MSAVI) generated the highest coefficient of determination (R2 = 0.93). MTVI1 and triangular VI had the highest second and third coefficient of determination with N content (R2 = 0.926 and 0.923, respectively). The classification accuracy assessment of the developed model was evaluated using several statistical parameters such as the independent t-test, and p-value. The accuracy assessment of the developed model was more than 77%.
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    The Effect of Diet and Farm Management on N2O Emissions from Dairy Farms Estimated from Farm Data
    (Basel : MDPI, 2021) Menardo, Simona; Lanza, Giacomo; Berg, Werner
    The N2O emissions of 21 dairy farms in Germany were evaluated to determine the feasi-bility of an estimation of emissions from farm data and the effects of the farm management, along with possible mitigation strategies. Emissions due to the application of different fertilisers, manure storage and grazing were calculated based on equations from the IPCC (Intergovernmental Panel of Climate Change) and German emission inventory. The dependence of the N2O emissions on fertiliser type and quantity, cultivated crops and diet composition was assessed via correlation analysis and linear regression. The N2O emissions ranged between 0.11 and 0.29 kg CO2eq per kilogram energy-corrected milk, with on average 60% resulting from fertilisation and less than 30% from fertiliser storage and field applications. The total emissions had a high dependence on the diet composition; in particular, on the grass/maize ratio and the protein content of the animal diet, as well as from the manure management. A linear model for the prediction of the N2O emissions based on the diet composition and the fertilisation reached a predictive power of R2 = 0.89. As a possible mitigation strategy, the substitution of slurry for solid manure would reduce N2O emissions by 40%. Feeding cows maize-based diets instead of grass-based diets could reduce them by 14%. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
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    Characterization of Bathyarchaeota genomes assembled from metagenomes of biofilms residing in mesophilic and thermophilic biogas reactors
    (London : BioMed Central Ltd., 2018) Maus, I.; Rumming, M.; Bergmann, I.; Heeg, K.; Pohl, M.; Nettmann, E.; Jaenicke, S.; Blom, J.; Pühler, A.; Schlüter, A.; Sczyrba, A.; Klocke, M.
    Background: Previous studies on the Miscellaneous Crenarchaeota Group, recently assigned to the novel archaeal phylum Bathyarchaeota, reported on the dominance of these Archaea within the anaerobic carbohydrate cycle performed by the deep marine biosphere. For the first time, members of this phylum were identified also in mesophilic and thermophilic biogas-forming biofilms and characterized in detail. Results: Metagenome shotgun libraries of biofilm microbiomes were sequenced using the Illumina MiSeq system. Taxonomic classification revealed that between 0.1 and 2% of all classified sequences were assigned to Bathyarchaeota. Individual metagenome assemblies followed by genome binning resulted in the reconstruction of five metagenome-assembled genomes (MAGs) of Bathyarchaeota. MAGs were estimated to be 65-92% complete, ranging in their genome sizes from 1.1 to 2.0 Mb. Phylogenetic classification based on core gene sets confirmed their placement within the phylum Bathyarchaeota clustering as a separate group diverging from most of the recently known Bathyarchaeota clusters. The genetic repertoire of these MAGs indicated an energy metabolism based on carbohydrate and amino acid fermentation featuring the potential for extracellular hydrolysis of cellulose, cellobiose as well as proteins. In addition, corresponding transporter systems were identified. Furthermore, genes encoding enzymes for the utilization of carbon monoxide and/or carbon dioxide via the Wood-Ljungdahl pathway were detected. Conclusions: For the members of Bathyarchaeota detected in the biofilm microbiomes, a hydrolytic lifestyle is proposed. This is the first study indicating that Bathyarchaeota members contribute presumably to hydrolysis and subsequent fermentation of organic substrates within biotechnological biogas production processes.
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    Limited life cycle andcost assessment for the bioconversion of lignin‐derived aromatics into adipic acid
    (New York, NY [u.a.] : Wiley, 2020) van Duuren, Jozef B.J.H.; de Wild, Paul J.; Starck, Sören; Bradtmöller, Christian; Selzer, Mirjam; Mehlmann, Kerstin; Schneider, Roland; Kohlstedt, Michael; Poblete‐Castr, Ignacio; Stolzenberger, Jessica; Barton, Nadja; Fritz, Michel; Scholl, Stephan; Venus, Joachim; Wittmann, Christoph
    Lignin is an abundant and heterogeneous waste byproduct of the cellulosic industry, which has the potential of being transformed into valuable biochemicals via microbial fermentation. In this study, we applied a fast-pyrolysis process using softwood lignin resulting in a two-phase bio-oil containing monomeric and oligomeric aromatics without syringol. We demonstrated that an additional hydrodeoxygenation step within the process leads to an enhanced thermochemical conversion of guaiacol into catechol and phenol. After steam bath distillation, Pseudomonas putida KT2440-BN6 achieved a percent yield of cis, cis-muconic acid of up to 95 mol% from catechol derived from the aqueous phase. We next established a downstream process for purifying cis, cis-muconic acid (39.9 g/L) produced in a 42.5 L fermenter using glucose and benzoate as carbon substrates. On the basis of the obtained values for each unit operation of the empirical processes, we next performed a limited life cycle and cost analysis of an integrated biotechnological and chemical process for producing adipic acid and then compared it with the conventional petrochemical route. The simulated scenarios estimate that by attaining a mixture of catechol, phenol, cresol, and guaiacol (1:0.34:0.18:0, mol ratio), a titer of 62.5 (g/L) cis, cis-muconic acid in the bioreactor, and a controlled cooling of pyrolysis gases to concentrate monomeric aromatics in the aqueous phase, the bio-based route results in a reduction of CO2-eq emission by 58% and energy demand by 23% with a contribution margin for the aqueous phase of up to 88.05 euro/ton. We conclude that the bio-based production of adipic acid from softwood lignins brings environmental benefits over the petrochemical procedure and is cost-effective at an industrial scale. Further research is essential to achieve the proposed cis, cis-muconic acid yield from true lignin-derived aromatics using whole-cell biocatalysts. © 2020 Wiley Periodicals, Inc.