Mikrobiologie der Fermentation in NawaRo-Biogasanlagen unter besonderer Berücksichtigung des Abbaus von Propionsäure

In NawaRo-Biogasanlagen (BGA) kann es durch das Angebot an leicht fermentierbaren Kohlenstoff¬quel¬len zu einer bakteriell bedingten Übersäuerung durch unerwünschte kurzkettige Fettsäuren kommen. Häufiger kommt es zur Akkumulation von Propionsäure. Methanogene Archaea können bei niedrigen pH-Werten nicht mehr wachsen. Somit kann der gesamte Prozess der mikrobiellen Bildung von Biogas zum Erliegen kom¬men, was für die Biogasbetreiber zu erheblichen finanziellen Verlusten führt. Das Ziel dieser Disserta¬tion war die Aufklärung der anaeroben bakteriellen Population, die in Biogasanlagen Propionsäure ab¬bauen kann. Aus Propionat entsteht dabei Acetat und Wasserstoff. Da dieser anaerobe Prozess endergon verläuft, kann Propionsäure anaerob nur abgebaut werden, wenn der Wasserstoffpartialdruck niedrig ge¬halten wird. Diese Aufgabe erfüllen in Biogasanalgen methanogene Archaea. Die sog. sekundären Gärer leben somit in synthropher Kultur mit methanogenen Archaea.rnIn dieser Arbeit wurden die Mikroorganismen von Propionsäure-abbauenden Anreicherungskulturen aus vier NawaRo-BGA‘s identifiziert und ihr Substrat- und Produktspektrum analysiert. Die Anreicherungskul¬turen wurden vom Prüf- und Forschungsinstitut e. V. in Pirmasens zur Verfügung gestellt. Durch Analyse der bakteriellen 16S rDNA-Sequenzen der erhaltenen stabilen Propionsäure-abbauenden Mischkulturen wurde gezeigt, dass sich unter den Bakterien hauptsächlich Verwandte von den Clostridiales, aber auch Bacteroides sp., δ-, ε- so¬wie γ-Proteobakterien, Spirochäten, Synergistales und ungewöhnlicher Weise auch Thermotogales befanden. Aus Propionsäure-abbauenden Mischkulturen und aus Fermentern mesophiler NawaRo-Biogasanlagen wurden anaerobe Bakterien und methanogene Archaea angereichert und isoliert. Es wurden aus den Propionsäure-abbauenden Mischkulturen Stämme in Reinkultur erhalten, die entsprechend der 16S rDNA-Analyse als Clostridium sartagoforme Stamm Ap1a520 und Proteiniphilum acetatigenes Stamm Fp1a520 identifiziert wurden. Sowohl aus Fermentern und Nachgärern von drei NawaRo-BGA‘s als auch aus zwei Laborfermentern des Leibniz-Instituts für Agrartechnik in Potsdam-Bornim e.V. (ATB) wurden Reinkulturen von methanogenen Archaea erhalten. Diese konnten den Species Methanobacterium formicicum, Metha¬noculleus bourgensis, Methanosarcina barkeri, Methanosarcina mazei, Methanosarcina sp., Methanosaeta concilii und Methanomethylovorans sp. zugeordnet werden. Damit wurden in dieser Arbeit unter anderem die typischen bisher nur durch molekularbiologische Methoden identifizierten Species methanogener Ar¬chaea aus unterschiedlichen Fermentern in Reinkultur erhalten. Dabei wurde gezeigt, dass die specifically amplified polymorphic DNA-PCR (SAPD-PCR) eine geeignete Methode darstellt, Stämme der gleichen Art methanogener Archaea voneinander zu unterscheiden. Die Methanproduktion der kultivierten methanoge¬nen Archaea wurde gaschromatographisch analysiert. Es zeigte sich, dass die hydrogenotrophe Metha¬nogenese der effizientere und ergiebigere Weg zur Bildung von Methan ist. Mit der Bestimmung der Zellzahl des Isolates Methanoculleus bourgensis Stamm TAF1.1 bei gleichzeitiger Messung der Methanbildung wurde gezeigt, dass die Methanbildung nicht zwangsläufig mit dem Wachstum korreliert. Ne-ben Pflanzenfasern beinhalteten das hergestellte Reaktorfiltrat in den Kultivierungsansätzen Acetat, die essentielle Aminosäure Valin und den Zuckeralkohol Glycerol. Gezielte Misch¬kul¬turen von sekundären Gärern mit methanogenen Isolaten ergaben einen fördernden Einfluss auf diese Bak¬terien durch hydrogenotrophe Archaea. Diese Bakterien bauten Substrate ab oder bildeten Produkte, die sie unter den gegebenen Bedingungen ohne hydrogenotrophe Archaea nicht umsetzen konnten.

The ethanolic fermentation pathway supports respiration and lipid biosynthesis in tobacco pollen

Rapid pollen tube growth requires a high rate of sugar metabolism to meet energetic and biosynthetic demands. Previous work on pollen sugar metabolism showed that tobacco pollen carry out efficient ethanolic fermentation concomitantly with a high rate of respiration (Bucher et al ., 1995). Here we show that the products of fermentation, acetaldehyde and ethanol, are further metabolised in a pathway that bypasses mitochondrial PDH. The enzymes involved in this pathway are pyruvate decarboxylase, aldehyde dehydrogenase and acetyl-CoA synthetase. Radiolabelling experiments show that during tobacco pollen tube growth label of C-14-ethanol is incorporated into CO2 as well as into lipids and other higher molecular weight compounds. A role for the glyoxylate cycle appears unlikely since activity of malate synthase, a key enzyme of the glyoxylate cycle, could not be detected.

Ethanolic fermentation: new functions for an old pathway

Ethanolic fermentation is an ancient metabolic pathway. In plants, it is a major route of {ATP} production under anaerobic conditions. In addition, recent developments suggest that the pathway has important functions in the presence of oxygen. Both of the enzymes required for the production of acetaldehyde and ethanol, pyruvate decarboxylase and alcohol dehydrogenase, are highly abundant in pollen, resulting in fermentation in fully oxygenated cells. Acetaldehyde toxicity is an inevitable side effect of aerobic fermentation. Could acetaldehyde be the elusive pollen factor that contributes to male sterility in cmsT maize? The versatility of this ancient pathway is also illustrated by the induction of aerobic fermentation by environmental stress and activation of a defense response by overexpression of pyruvate decarboxylase.

Proceedings of the 10th Annual Biochemical Engineering Symposium

This the tenth in a series of symposia devoted to talks by students on their biochemical engineering research. The first, third, fifth, and ninth were at Kansas State University in Manhattan, the second and fourth were at the University of Nebraska–Lincoln, the sixth was in Kansas City in conjunction with the 81st American Institute of Chemical Engineers National Meeting, the seventh was at Iowa State University in Ames, and the eighth was held at the University of Missouri–Columbia. Contents"Combined Autohydrolysis-Organosolv Pretreatment of Lignocellulosic Materials," Robert A. Lewis, Colorado State University "An Investigation of Cellulase Activity Assays," Minhhuong Nguyen, University of Missouri–Columbia "Action Pattern of a Xylobiohydrolase from Aspergillus niger," Mary M. Frederick, Iowa State University "Estimation of Heats of Combustion of Biomass from Elemental Analysis Using Available Electron Concepts," Snehal A. Patel, Kansas State University "Design of a Wheat Straw to Ethanol Conversion Facility," Michael M. Meagher, Colorado State University "Effects of Salt, Heat, and Physical Form on the Fermentation of Bananas," Carl Drewel, University of Missouri–Columbia "Gas Hold-up in the Downflow Section of a Split Cylinder Airlift Column," Vasanti Deshpande, Kansas State University "Measurement of Michaelis Constants for Soluble and Immobilized Glucoamylase," Robert A. Lesch, Iowa State University "Kinetics of Alkaline Oxidation and Degradation of Sugars," Alfred R. Fratzke, Iowa State University "Stability of Cereal Protein During Microbial Growth on Grain Dust," Bamidele O. Solomon, Kansas State University

Concentration profiles in batch fermentation of hydrolyzed lignocellulose

Experimental data was obtained for profiling changes in concentrations of two inhibiting compounds in batch fermentation of a synthesized liquor resembling hydrolyzed lignocellulose, a furan (furfural) and a phenolic compound (vanillin), along with standard fermentation data, i.e. substrate, biomass and ethanol concentrations. The initial inhibitor concentrations and fermentation temperatures in the 18 experiments were varied according to a two-level complete center composite experimental design. Based upon these observed variations in the fermentative behavior, the fermentation kinetics were modeled, as published in the corresponding article, including microbial conversion rates of the inhibitive compounds into their less toxic derivatives.

Impacts of projected maximum temperature extremes for C21 by an ensemble of regional climate models on cereal cropping systems in the Iberian Peninsula

Crops growing in the Iberian Peninsula may be subjected to damagingly high temperatures during the sensitive development periods of flowering and grain filling. Such episodes are considered important hazards and farmers may take insurance to offset their impact. Increases in value and frequency of maximum temperature have been observed in the Iberian Peninsula during the 20th century, and studies on climate change indicate the possibility of further increase by the end of the 21st century. Here, impacts of current and future high temperatures on cereal cropping systems of the Iberian Peninsula are evaluated, focusing on vulnerable development periods of winter and summer crops. Climate change scenarios obtained from an ensemble of ten Regional Climate Models (multimodel ensemble) combined with crop simulation models were used for this purpose and related uncertainty was estimated. Results reveal that higher extremes of maximum temperature represent a threat to summer-grown but not to winter-grown crops in the Iberian Peninsula. The study highlights the different vulnerability of crops in the two growing seasons and the need to account for changes in extreme temperatures in developing adaptations in cereal cropping systems. Finally, this work contributes to clarifying the causes of high-uncertainty impact projections from previous studies.

Effects of the main cereal and type of fat of the diet on productive performance and egg quality of brown egg laying hens from twenty two to fifty four weeks of age

The influence of the main cereal and supplemental fat of the diet on productive performance and egg quality was studied in 756 brown-egg laying hens from 22 to 54 weeks of age. The experiment was conducted as a completely randomized design with 9 treatments arranged factorially with 3 cereals (dented corn, soft wheat, and barley) and 3 types of fat [soy oil (SBO), acidulated soapstocks (AOS), and lard].

Effecto del cereal principal y el tipo de grasa en la dieta sobre los parámetros productivos en gallinas rubias en el periodo

Maíz, trigo y cebada son cereales comúnmente utilizados en piensos para ponedoras. A nivel práctico estos piensos incluyen un mínimo de 25 a 30% de maíz para asegurar altos consumos y mejorar el tamaño del huevo. Esta práctica puede estar relacionada con la mejora de la estructura del pienso maíz, así como al mayor contenido en ácido linoléico (LIN) de este cereal, que puede dar lugar en un incremento del peso del huevo, especialmente cuando el consumo es bajo. La adición de grasa al pienso aumenta la concentración energética del mismo y en general, el contenido en LIN (Schutze and Jensen, 1963). En este trabajo se investigó el efecto del tipo de cereal y grasa añadida sobre la productividad en ponedoras rubias. Un segundo objetivo fue evaluar la relación entre contenido en LIN del pienso y el tamaño del huevo.

Zonificación del espacio por agrupación de unidades superficiales. Aplicación a la gestión de sistemas extensivos del tipo cereal-ovino

RESUMEN Los procesos de diseño de zonas o diseño del territorio implican la partición de un espacio geográfico, organizado en un conjunto de unidades de área, en diferentes regiones o zonas según un conjunto especifico de criterios que varían en función del campo de aplicación. En la mayoría de los casos, el objetivo fundamental consiste en crear zonas de tamaño aproximadamente igual respecto a uno o varios atributos de medida -de carácter cuantitativo- (zonas con igual número de habitantes, igual promedio de ventas...). Sin embargo, están apareciendo nuevas aplicaciones, algunas en el contexto de las políticas de desarrollo sostenible, cuya finalidad es la definición de regiones con un tamaño predeterminado, no necesariamente similar. Además, en estos casos las zonas han de formarse en torno a un conjunto específico de posiciones, semillas o generadores. Este tipo de particiones no han sido lo suficientemente investigadas, de manera que no se conocen modelos de solución para la delimitación automática de las zonas. En esta tesis se ha diseñado un nuevo método basado en una versión discreta del diagrama de Voronoi con peso aditivo adaptativo (DVPAA), que permite la partición de un espacio bidimensional en zonas de un tamaño específico, considerando tanto la posición como el peso de cada uno de los generadores. El método consiste en resolver repetidamente un tradicional diagrama de Voronoi con peso aditivo, de forma que los pesos de cada generador se actualizan en cada iteración. En el proceso de cálculo de distancias se usa una métrica basada en el camino más corto, lo que garantiza que la partición obtenida esté formada por un conjunto de zonas conexas. La heurística diseñada se integra en una aplicación prototipo, desarrollada en un entorno SIG (Sistemas de Información Geográfica), que permite el trazado automático de zonas según los criterios anteriormente expuestos. Para analizar la viabilidad del método se ha utilizado como caso de estudio la gestión de los recursos pastorales para la ganadería extensiva en tres municipios de Castilla-La Mancha. Las pruebas realizadas ponen de manifiesto que la heurística diseñada, adaptada a los criterios que se plantean en el contexto de la gestión de sistemas extensivos agropecuarios, es válida para resolver este tipo de problemas de partición. El método propuesto se caracteriza por su eficacia en el tratamiento de un gran número de unidades superficiales en formato vectorial, generando soluciones que convergen con relativa rapidez y verifican los criterios establecidos. En el caso estudiado, aunque la posición prefijada de los generadores reduce considerablemente la complejidad del problema, existen algunas configuraciones espaciales de estos elementos para las que el algoritmo no encuentra una solución satisfactoria, poniéndose de manifiesto una de las limitaciones de este modelo. Tal y como se ha podido comprobar, la localización de los generadores puede tener un considerable impacto en la zonificación resultante, por lo que, de acuerdo con Kalcsics et al. (2005), una selección "inadecuada" difícilmente puede generar regiones válidas que verifiquen los criterios establecidos. ABSTRACT Tenitory or zone design processes entail partitioning a geographic space, organized as a set of basic areal units, into different regions or zones according to a specific set of entena that are dependent on the application context. In most cases the aim is to create zones that have approximately equal sizes with respect to one or several measure attributes (zones with equal numbers of inhabitants, same average sales, etc). However, some of the new applications that have emerged, particularly in the context of sustainable development policies, are aimed at defining zones of a predetermined, though not necessarily similar, size. In addition, the zones should be built around a given set of positions, seeds or generators. This type of partitioning has not been sufñciently researched; therefore there are no known approaches for automated zone delimitation. This thesis proposes a new method based on a discrete versión of the Adaptive Additively Weighted Voronoi Diagram (AAWVD) that makes it possible to partition a 2D space into zones of specific sizes, taking both the position and the weight of each (seed) generator into account. The method consists of repeatedly solving a traditional additively weighted Voronoi diagram, so that the weights of each generator are updated at every iteration. The partition s zones are geographically connected nsing a metric based 011 the shortest path. The proposed heuristic lias been included in an application, developed in a GIS environment that allows the automated zone delimitation according to the mentioned criteria. The management of the extensive farming system of three municipalities of Castilla-La Mancha (Spain) has been used as study case to analyze the viability of the method. The tests carried out have established that the proposed method, adapted to the criteria of this application field, is valid for solving this type of partition problem. The applied algorithm is capable of handling a high number of vector areal units, generating solutions that converge in a reasonable CPU time and comply with the imposed constraints. Although the complexity of this problem is greatly reduced when the generator's positions are fixed, in many cases, these positions impose a spatial confignration that the algorithm proposed is unable to solve, thus revealing one of the limitations of this method. It has been shown that the location of the generators has a considerable impact on the final solution, so that, as Kalcsics et al. (2005) observed, an "inadequate" selection can hardly generate valid zones that comply with the established criteria.

Influence of the Main Cereal and Feed Form of the Diet on Performance and Digestive Tract Traits of Brown-Egg Laying Pullets.

The influence of the main cereal and feed form of the diet on performance and digestive tract traits was studied in 576 brown-egg laying pullets from 1 to 120 d of age. From 1 to 45 d of age, 4 diets arranged factorially with 2 cereals (corn vs. wheat) and 2 feed forms (mash vs. pellets) were used. Each treatment was replicated 6 times (24 pullets per replicate). From 46 to 120 d of age, all diets were offered in mash form and the only difference among diets was the cereal used. Cumulatively, pullets fed the corn diets had higher BW gain (P < 0.05) but similar feed conversion ratio as pullets fed the wheat diets. From 1 to 45 d of age, pullets fed pellets consumed more feed (P < 0.001) and had higher BW gain (P < 0.001) than those fed mash. Most of the beneficial effects of pelleting on productive performance were still evident at 120 d of age. At 45 d of age, gizzard weight (g/kg of BW) was higher (P < 0.01) in pullets fed corn than in pullets fed wheat diets. Feeding pellets reduced the relative weight of the digestive tract and the gizzard (P < 0.001) as well as the length (cm/kg of BW) of the small intestine (P < 0.01) at both ages. The pH of gizzard contents at 120 d of age was not affected by cereal but was lower in pullets that were fed mash from 1 to 45 d of age (P < 0.01). We conclude that wheat can be used in substitution of corn in pullet diets with only a slight reduction in BW gain and that feeding pellets from 1 to 45 d of age increased BW gain and pH of the gizzard and reduced the relative weight of the gizzard and the length of the gastrointestinal tract at 120 d of age.

Multiple avirulence paralogues in cereal powdery mildew fungi may contribute to parasite fitness and defeat of plant resistance

Powdery mildews, obligate biotrophic fungal parasites on a wide range of important crops, can be controlled by plant resistance (R) genes, but these are rapidly overcome by parasite mutants evading recognition. It is unknown how this rapid evolution occurs without apparent loss of parasite fitness. R proteins recognize avirulence (AVR) molecules from parasites in a gene-for-gene manner and trigger defense responses. We identify AVRa10 and AVRk1 of barley powdery mildew fungus, Blumeria graminis f sp hordei (Bgh), and show that they induce both cell death and naccessibility when transiently expressed in Mla10 and Mlk1 barley (Hordeum vulgare) varieties, respectively. In contrast with other reported fungal AVR genes, AVRa10 and AVRk1 encode proteins that lack secretion signal peptides and enhance infection success on susceptible host plant cells. AVRa10 and AVRk1 belong to a large family with mayor que30 paralogues in the genome of Bgh, and homologous sequences are present in other formae speciales of the fungus infecting other grasses. Our findings imply that the mildew fungus has a repertoire of AVR genes, which may function as effectors and contribute to parasite virulence. Multiple copies of related but distinct AVR effector paralogues might enable populations of Bgh to rapidly overcome host R genes while maintaining virulence.

Sum Insured Determination for Cereal, Citrus and Vineyards in the Spanish Agricultural Insurance System.

In general, insurance is a form of risk management used to hedge against a contingent loss. The conventional definition is the equitable transfer of a risk of loss from one entity to another in exchange for a premium or a guaranteed and quantifiable small loss to prevent a large and possibly devastating loss being agricultural insurance a special line of property insurance. Agriculture insurance, as actually are designed in the Spanish scenario, were established in 1978. At the macroeconomic insurance studies scale, it is necessary to know a basic element for the insurance actuarial components: sum insured. When a new risk assessment has to be evaluated in the insurance framework, it is essential to determinate venture capital in the total Spanish agriculture. In this study, three different crops (cereal, citrus and vineyards) cases are showed to determinate sum insured as they are representative of the cases found in the Spanish agriculture. Crop sum insured is calculated by the product of crop surface, unit surface production and crop price insured. In the cereal case, winter as spring cereal sowing, represents the highest Spanish crop surface, above to 6 millions of hectares (ha). Meanwhile, the four citrus species (oranges, mandarins, lemons and grapefruits) occupied an extension just over 275.000 ha. On the other hand, vineyard target to wine process shows almost one million of ha in Spain.