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    Effect of Cereal α-Amylase/Trypsin Inhibitors on Developmental Characteristics and Abundance of Digestive Enzymes of Mealworm Larvae (Tenebrio molitor L.)
    (Basel : MDPI, 2021) Sagu, Sorel Tchewonpi; Landgräber, Eva; Henkel, Ina M; Huschek, Gerd; Homann, Thomas; Bußler, Sara; Schlüter, Oliver K.; Rawel, Harshadrai
    The objective of this work was to investigate the potential effect of cereal α-amylase/trypsin inhibitors (ATIs) on growth parameters and selective digestive enzymes of Tenebrio molitor L. larvae. The approach consisted of feeding the larvae with wheat, sorghum and rice meals containing different levels and composition of α-amylase/trypsin inhibitors. The developmental and biochemical characteristics of the larvae were assessed over feeding periods of 5 h, 5 days and 10 days, and the relative abundance of α-amylase and selected proteases in larvae were determined using liquid chromatography tandem mass spectrometry. Overall, weight gains ranged from 21% to 42% after five days of feeding. The larval death rate significantly increased in all groups after 10 days of feeding (p < 0.05), whereas the pupation rate was about 25% among larvae fed with rice (Oryza sativa L.) and Siyazan/Esperya wheat meals, and only 8% and 14% among those fed with Damougari and S35 sorghum meals. As determined using the Lowry method, the protein contents of the sodium phosphate extracts ranged from 7.80 ± 0.09 to 9.42 ± 0.19 mg/mL and those of the ammonium bicarbonate/urea reached 19.78 ± 0.16 to 37.47 ± 1.38 mg/mL. The total protein contents of the larvae according to the Kjeldahl method ranged from 44.0 and 49.9 g/100 g. The relative abundance of α-amylase, CLIP domain-containing serine protease, modular serine protease zymogen and C1 family cathepsin significantly decreased in the larvae, whereas dipeptidylpeptidase I and chymotrypsin increased within the first hours after feeding (p < 0.05). Trypsin content was found to be constant independently of time or feed material. Finally, based on the results we obtained, it was difficult to substantively draw conclusions on the likely effects of meal ATI composition on larval developmental characteristics, but their effects on the digestive enzyme expression remain relevant.
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    Establishment of a Laboratory Scale Set-Up with Controlled Temperature and High Humidity to Investigate Dry Matter Losses of Wood Chips from Poplar during Storage
    (Basel : MDPI, 2022) Hernandez-Estrada, Albert; Pecenka, Ralf; Dumfort, Sabrina; Ascher-Jenull, Judith; Lenz, Hannes; Idler, Christine; Hoffmann, Thomas
    The aim of this work was to improve the understanding of dry matter losses (DML) that occur in wood chips during the initial phase of storage in outdoor piles. For this purpose, a laboratory scale storage chamber was developed and investigated regarding its ability to recreate the conditions that chips undergo during the initial phase of outdoor storage. Three trials with poplar Max-4 (Populus maximowiczii Henry  Populus nigra L.) chips were performed for 6–10 weeks in the storage chamber under controlled temperature and assisted humidity. Two different setups were investigated to maintain a high relative humidity (RH) inside the storage chamber; one using water containers, and one assisted with a humidifier. Moisture content (MC) and DML of the chips were measured at different storage times to evaluate their storage behaviour in the chamber. Additionally, microbiological analyses of the culturable fraction of saproxylic microbiota were performed, with a focus on mesophilic fungi, but discriminating also xerophilic fungi, and mesophilic bacteria, with focus on actinobacteria, in two trials, to gain a view on the poplar wood chip-inhabiting microorganisms as a function of storage conditions (moisture, temperature) and time. Results show that DML up to 8.8–13.7% occurred in the chips within 6–10 storage weeks. The maximum DML were reached in the trial using the humidifier, which seemed a suitable technique to keep a high RH in the testing chamber, and thus, to analyse the wood chips in conditions comparable to those in outdoor piles during the initial storage phase.
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    Integration of Solid State and Submerged Fermentations for the Valorization of Organic Municipal Solid Waste
    (Basel : MDPI, 2021) Martău, Gheorghe-Adrian; Unger, Peter; Schneider, Roland; Venus, Joachim; Vodnar, Dan Cristian; López-Gómez, José Pablo
    Solid state fermentation (SsF) is recognized as a suitable process for the production of enzymes using organic residues as substrates. However, only a few studies have integrated an evaluation of the feasibility of applying enzymes produced by SsF into subsequent hydrolyses followed by the production of target compounds, e.g., lactic acid (LA), through submerged-liquid fermentations (SmF). In this study, wheat bran (WB) was used as the substrate for the production of enzymes via SsF by Aspergillus awamori DSM No. 63272. Following optimization, cellulase and glucoamylase activities were 73.63 ± 5.47 FPU/gds and 107.10 ± 2.63 U/gdb after 7 days and 5 days of fermentation, respectively. Enzymes were then used for the hydrolysis of the organic fraction of municipal solid waste (OFMSW). During hydrolysis, glucose increased considerably with a final value of 19.77 ± 1.56 g/L. Subsequently, hydrolysates were fermented in SmF by Bacillus coagulans A166 increasing the LA concentration by 15.59 g/L. The data reported in this study provides an example of how SsF and SmF technologies can be combined for the valorization of WB and OFMSW.
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    Pilot Scale for Production and Purification of Lactic Acid from Ceratonia siliqua L. (Carob) Bagasse
    (Basel : MDPI, 2022) Azaizeh, Hassan; Abu Tayeh, Hiba Nazmi; Schneider, Roland; Venus, Joachim
    The bioconversion of lignocellulose and organic waste bagasse to lactic acid (LA) is an important alternative process requiring valorization as a potentially viable method in the production of pure LA, to be utilized for various purposes. Carob (Ceratonia siliqua L.) biomass was used for the production of LA, using a thermophilic Bacillus coagulans isolate, cultivated in a batch pilot scale of 35 L fermenters without yeast extract supplementation, and operated for 50 h. During the fermentation process, most of the degradable sugar was consumed within 35 h and resulted in the production of 46.9 g/L LA, with a calculated LA yield of 0.72 g/g sugars and productivity at the log phase of 1.69 g/L/h. The use of LA for different industrial applications requires high purity; therefore, a downstream process (DSP) consisting of different purification stages was used, enabling us to reach up to 99.9% (w/w) product purity, which indicates that the process was very effective. The overall almost pure L-LA yield of the DSP was 56%, which indicates that a considerable amount of LA (46%) was lost during the different DSP stages. This is the first study in which carob biomass bagasse has been tested on a pilot scale for LA production, showing the industrial feasibility of the fermentation process.
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    The Role of Streptococcus spp. in Bovine Mastitis
    (Basel : MDPI, 2021) Kabelitz, Tina; Aubry, Etienne; van Vorst, Kira; Amon, Thomas; Fulde, Marcus
    The Streptococcus genus belongs to one of the major pathogen groups inducing bovine mastitis. In the dairy industry, mastitis is the most common and costly disease. It not only negatively impacts economic profit due to milk losses and therapy costs, but it is an important animal health and welfare issue as well. This review describes a classification, reservoirs, and frequencies of the most relevant Streptococcus species inducing bovine mastitis (S. agalactiae, S. dysgalactiae and S. uberis). Host and environmental factors influencing mastitis susceptibility and infection rates will be discussed, because it has been indicated that Streptococcus herd prevalence is much higher than mastitis rates. After infection, we report the sequence of cow immune reactions and differences in virulence factors of the main Streptococcus species. Different mastitis detection techniques together with possible conventional and alternative therapies are described. The standard approach treating streptococcal mastitis is the application of ß-lactam antibiotics. In streptococci, increased antimicrobial resistance rates were identified against enrofloxacin, tetracycline, and erythromycin. At the end, control and prevention measures will be considered, including vaccination, hygiene plan, and further interventions. It is the aim of this review to estimate the contribution and to provide detailed knowledge about the role of the Streptococcus genus in bovine mastitis.
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    Optical Spectrometry to Determine Nutrient Concentrations and other Physicochemical Parameters in Liquid Organic Manures: A Review
    (Basel : MDPI, 2022) Horf, Michael; Vogel, Sebastian; Drücker, Harm; Gebbers, Robin; Olfs, Hans-Werner
    Nutrient concentrations in livestock manures and biogas digestates show a huge variability due to disparities in animal husbandry systems concerning animal species, feed composition, etc. Therefore, a nutrient estimation based on recommendation tables is not reliable when the exact chemical composition is needed. The alternative, to analyse representative fertilizer samples in a standard laboratory, is too time-and cost-intensive to be an accepted routine method for farmers. However, precise knowledge about the actual nutrient concentrations in liquid organic fertilizers is a prerequisite to ensure optimal nutrient supply for growing crops and on the other hand to avoid environmental problems caused by overfertilization. Therefore, spectrometric methods receive increasing attention as fast and low-cost alternatives. This review summarizes the present state of research based on optical spectrometry used at laboratory and field scale for predicting several parameters of liquid organic manures. It emphasizes three categories: (1) physicochemical parameters, e.g., dry matter, pH, and electrical conductivity; (2) main plant nutrients, i.e., total nitrogen, ammonium nitrogen, phosphorus, potassium, magnesium, calcium, and sulfur; and (3) micronutrients, i.e., manganese, iron, copper, and zinc. Furthermore, the commonly used sample preparation techniques, spectrometer types, measuring modes, and chemometric methods are presented. The primarily promising scientific results of the last 30 years contributed to the fact that near-infrared spectrometry (NIRS) was established in commercial laboratories as an alternative method to wet chemical standard methods. Furthermore, companies developed technical setups using NIRS for on-line applications of liquid organic manures. Thus, NIRS seems to have evolved to a competitive measurement procedure, although parts of this technique still need to be improved to ensure sufficient accuracy, especially in quality management.
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    Peptaibol Production and Characterization from Trichoderma asperellum and Their Action as Biofungicide
    (Basel : MDPI, 2022) Alfaro-Vargas, Pamela; Bastos-Salas, Alisson; Muñoz-Arrieta, Rodrigo; Pereira-Reyes, Reinaldo; Redondo-Solano, Mauricio; Fernández, Julián; Mora-Villalobos, Aníbal; López-Gómez, José Pablo
    Peptaibols (Paib), are a class of biologically active peptides isolated from soil, fungi and molds, which have interesting properties as antimicrobial agents. Paib production was optimized in flasks by adding sucrose as a carbon source, 2-aminoisobutyric acid (Aib) as an additive amino acid, and F. oxysporum cell debris as an elicitor. Paib were purified, sequenced and identified by High-performance liquid chromatography (HPLC)coupled to mass spectrometry. Afterward, a Paib extract was obtained from the optimized fermentations. The biological activity of these extracts was evaluated using in vitro and in vivo methods. The extract inhibited the growth of specific plant pathogens, and it showed inhibition rates similar to those from commercially available fungicides. Growth inhibition rates were 92.2, 74.2, 58.4 and 36.2% against Colletotrichum gloeosporioides, Botrytis cinerea, Alternaria alternata and Fusarium oxysporum, respectively. Furthermore, the antifungal activity was tested in tomatoes inoculated with A. alternata, the incidence of the disease in tomatoes treated with the extract was 0%, while the untreated fruit showed a 92.5% incidence of infection Scanning electron microscopy images showed structural differences between the fungi treated with or without Paib. The most visual alterations were sunk and shriveled morphology in spores, while the hyphae appeared to be fractured, rough and dehydrated.
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    Indicative Marker Microbiome Structures Deduced from the Taxonomic Inventory of 67 Full-Scale Anaerobic Digesters of 49 Agricultural Biogas Plants
    (Basel : MDPI, 2021) Hassa, Julia; Klang, Johanna; Benndorf, Dirk; Pohl, Marcel; Hülsemann, Benedikt; Mächtig, Torsten; Effenberger, Mathias; Pühler, Alfred; Schlüter, Andreas; Theuerl, Susanne
    There are almost 9500 biogas plants in Germany, which are predominantly operated with energy crops and residues from livestock husbandry over the last two decades. In the future, biogas plants must be enabled to use a much broader range of input materials in a flexible and demand-oriented manner. Hence, the microbial communities will be exposed to frequently varying process conditions, while an overall stable process must be ensured. To accompany this transition, there is the need to better understand how biogas microbiomes respond to management measures and how these responses affect the process efficiency. Therefore, 67 microbiomes originating from 49 agricultural, full-scale biogas plants were taxonomically investigated by 16S rRNA gene amplicon sequencing. These microbiomes were separated into three distinct clusters and one group of outliers, which are characterized by a specific distribution of 253 indicative taxa and their relative abundances. These indicative taxa seem to be adapted to specific process conditions which result from a different biogas plant operation. Based on these results, it seems to be possible to deduce/assess the general process condition of a biogas digester based solely on the microbiome structure, in particular on the distribution of specific indicative taxa, and without knowing the corresponding operational and chemical process parameters. Perspectively, this could allow the development of detection systems and advanced process models considering the microbial diversity.
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    UAV Oblique Imagery with an Adaptive Micro-Terrain Model for Estimation of Leaf Area Index and Height of Maize Canopy from 3D Point Clouds
    (Basel : MDPI, 2022) Li, Minhui; Shamshiri, Redmond R.; Schirrmann, Michael; Weltzien, Cornelia; Shafian, Sanaz; Laursen, Morten Stigaard
    Leaf area index (LAI) and height are two critical measures of maize crops that are used in ecophysiological and morphological studies for growth evaluation, health assessment, and yield prediction. However, mapping spatial and temporal variability of LAI in fields using handheld tools and traditional techniques is a tedious and costly pointwise operation that provides information only within limited areas. The objective of this study was to evaluate the reliability of mapping LAI and height of maize canopy from 3D point clouds generated from UAV oblique imagery with the adaptive micro-terrain model. The experiment was carried out in a field planted with three cultivars having different canopy shapes and four replicates covering a total area of 48 × 36 m. RGB images in nadir and oblique view were acquired from the maize field at six different time slots during the growing season. Images were processed by Agisoft Metashape to generate 3D point clouds using the structure from motion method and were later processed by MATLAB to obtain clean canopy structure, including height and density. The LAI was estimated by a multivariate linear regression model using crop canopy descriptors derived from the 3D point cloud, which account for height and leaf density distribution along the canopy height. A simulation analysis based on the Sine function effectively demonstrated the micro-terrain model from point clouds. For the ground truth data, a randomized block design with 24 sample areas was used to manually measure LAI, height, N-pen data, and yield during the growing season. It was found that canopy height data from the 3D point clouds has a relatively strong correlation (R2 = 0.89, 0.86, 0.78) with the manual measurement for three cultivars with CH90 . The proposed methodology allows a cost-effective high-resolution mapping of in-field LAI index extraction through UAV 3D data to be used as an alternative to the conventional LAI assessments even in inaccessible regions.
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    Readiness for Innovation of Emerging Grass-Based Businesses
    (Basel : MDPI, 2022) Orozco, Richard; Grundmann, Philipp
    New business opportunities based on grassland and green fodder present a promising avenue to realize the transition towards a circular and sustainable bio-based economy. Yet, such potential remains largely untapped and grass-based products and businesses remain a small niche in the global economy. To understand this phenomenon, this paper introduces and operationalizes a model to assess innovation readiness built around seven focus areas: technology, manufacturing, business, IPR, customer, team, and funding readiness with their own detailed “progress scales.” We employ necessary condition analysis (NCA) to identify limiting factors and bottlenecks in actual business situations. Our results reveal that lack of consumer awareness, infant conversion technologies and paucity of long-term investments that support emerging bio-based businesses are the most limiting conditions for the growth of emerging grass-based markets. The present study advances our understanding of the factors that limit complex innovations in grassland systems. Focusing on necessary conditions in a coordinated way between practitioners and policy makers by giving priority to fostering positive awareness of bioeconomy businesses, developing conversion technologies, and improving access to capital is a recommended approach to foster emerging grass-based innovations.