Browsing by Subject "Metabolism"
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Publication Effects of strain, lifespan and dietary myo-inositol sources on poultry metabolism(2020) Gonzalez Uarquin, Duvan Fernando; Huber, KorinnaPoultry production has shown a significant increase during the last decade. Meat and egg industry rapid growth implicates accelerating metabolic rate and general performance of birds. To maintain a high level of production, several strategies to achieve optimal raising and feeding have been implemented. Previous studies demonstrated the importance of MI metabolism on animal physiology; however, at present there is a substantial lack of information about the roles of MI and its metabolism in poultry. For instance, no information is available about MI concentration in organs of poultry. Moreover, it remains no elucidated, which are the effects of dietary sources of MI such as dietary phytase or pure MI supplementation. This thesis focused on gaining a comprehensive understanding of the potential roles of strain, productive period, and dietary sources of MI on poultry metabolism. To obtain the state of the art research on MI metabolism and its dietary sources in poultry, a comprehensive review of dietary MI was written (manuscript 1, chapter 3). This review revised information about MI in poultry such as feed sources, transport and cell metabolism, physiological meaning, and the influence of dietary MI in poultry. The revision indicated that MI appears to play critical roles in several different metabolic pathways so that understanding them could be an essential approach for future research in poultry. The second study was performed to study the effects of phytase and pure MI supplementation on the metabolite profile of broilers (manuscript 2, chapter 3). It was observed that phytase supplementation affected differently the metabolite profile than the supplementation of pure MI. Metabolites affected by phytase comprised several groups of metabolites such as acylcarnitines, phosphatidylcholines, sphingomyelins, lysophosphatidylcholines, and biogenic amines, whereas pure MI supplementation increased plasma concentrations of dopamine and serotonin. The third study was performed to get preliminary information about the effects caused by dietary phytase on systemic MI on the gastrointestinal tract, blood, and organs MI of broiler chickens (manuscript 3, chapter 3). Supplementation of 1500 FTU phytase/kg feed increased plasma and kidney MI concentrations. Plasma MI correlated negatively with InsP6 and positively with intestinal MI concentrations. A fourth study gave a general description of MI concentrations and general metabolite profile during the productive life of Lohmann Classic Brown and Lohmann LSL Classic laying hens. It was found that productive period affected MI and MI key enzymes expression. Moreover, the analyses showed differences in metabolite profiles being the onset of egg production, a determinant point. Differences were attributed to different groups of metabolites such as amino acids, biogenic amines, phosphatidylcholines, lysophosphatidylcholines, and sphingomyelins. The above mentioned, indicated each strain could express different MI concentrations and metabolite profiles during distinct productive periods what should be considered to future interventions. To conclude, findings from these investigations suggested intrinsic traits such as breed and stage of production and diet could affect MI and MI key enzymes expression as well as metabolite profiles. Future studies are needed to establish the roles of MI on poultry metabolism.Publication Escherichia coli O157:H7 und seine Adaption an verschiedene Wachstumsbedingungen in vitro : Untersuchung des intrazellulären Proteoms und Kohlenhydratmetabolismus(2014) Polzin, Sabrina; Schmidt, HerbertThe aim of this study was to investigate the linkage of metabolism and pathogenicity of EHEC bacteria under certain environmental conditions in vitro. During the investigations the influence of environmental conditions could be shown on the protein as well as on the gene expression level. In experiments using the culturel media SIEM and SCEM compared to TSB as a reference, differential expression of proteins and genes were shown. For example, decreasing motility and the formation of biofilm-like substances were observed. Furthermore, differences in metabolism and expression of chaperons and stress response systems were detected. After culturing EHEC bacteria in experimental media, increased glucose and pyruvate metabolism, increased amino acid biosynthesis, as well as increased chaperon levels were found, in contrast to decreased expression levels of stress associated proteins. The experimental procedure also affected the presence of pathogenicity factors, like EspP and EspB. Notably, the environmental conditions induced the glutamic acid resistance, providing the EHEC bacteria an unhindered passage through the human gastrointestinal tract, especially the acidic environments like the stomach. Additionally, the sialic acid esterase Z1466 along with shiga toxin, was strongly expressed. This could provide an advantage over the intestinal micro flora, in respect of growth. Effects onto glucose metabolism and amino acid biosynthesis were investigated by isotopoloque profiling experiments, prepared thankworthy by Dr. rer. nat. Wolfgang Eisenreich (Munich). To conclude, stress response and expression of pathogenicity factors could be induced in vitro in the cultural media elected, and without the need of a human host. The experiment also advices the complex regulation of different processes in bacterial cells as response to external factors.Publication Genome comparison of ‘Candidatus Phytoplasma rubi’ with genomes of other 16SrV phytoplasmas highlights special group features(2023) Böhm, Jan Werner; Duckeck, Dominik; Duduk, Bojan; Schneider, Bernd; Kube, MichaelPhytoplasmas are associated with important bacterial diseases, causing severe symptoms in agricultural and ornamental crops. ‘Candidatus Phytoplasma rubi’, associated with the Rubus stunt in raspberries (Rubus idaeus) and blackberries (Rubus subgenus Rubus), causes shortened internodes, elongated sepals, proliferation, phyllody, and virescence. The recently published genome of ‘Ca. P. rubi’ RS enabled a comprehensive genomic comparison to the complete genomes of 16SrV phytoplasmas, comprising strains of the flavescence dorée-associated phytoplasma CH and two ‘Candidatus Phytoplasma ziziphi’ strains. Besides the typical transporters and metabolic features of phytoplasmas, the phosphorolysis of sucrose and the utilization of the carboxylic acid L-lactate became apparent for the 16SrV-group. With respect to the effector repertoire and the encoded immunodominant membrane proteins involved in host colonization, the group revealed conserved features that comprise the variable membrane proteins A and B. However, SAP11- and SAP54 orthologs were limited to ‘Ca. P. rubi’ RS and ‘Ca. P. ziziphi’. Genome-sequence-based phylogenetic analysis supports the close relationship of these genomes relative to alder yellows phytoplasmas. The analyses supported the impact of the mobilome on phytoplasma evolution but also highlighted that there is the possibility of identifying phytoplasmas with a larger metabolic repertoire in the future.Publication Optimizing pome fruits storage(2024) Balkees, Basem Mahmoud; Zörb, ChristianHorticultural perishables are prone to fast deterioration and high losses. As the volume of production increases, further challenges are imposed to preserve these valuable foods in terms of quantity and quality. Accelerated development in science and applied technologies has helped to radically prolong the useful post-harvest life of horticultural produce. Notable success has been achieved with apples and pears as a result of their comparatively higher storability. Effective procedures and techniques have been developed to handle these fruits, including pre- and post-harvest physical and chemical treatments, in conjunction with the cutting-edge storage systems. However, certain procedures cannot be universally applicable. Considerations such as genotypic differences, climatic variations, and production factors necessitate adaptation of post-harvest practices to accommodate changing variables. This thesis describes the findings of experiments conducted as a contribution to advance preservation practices for apples and pears. Furthermore, we sought to investigate the biological actions underlying changes in fruit quality during storage and how these were influenced by various treatments. The first experiment was conducted to preserve the quality of late-harvested ‘Galaxy’ apples during extended storage. The effects of 1-methylcyclopropene (1 MCP) treatment and storage conditions on postharvest quality were analysed. Alongside quality measures, indicators like ethylene production, respiration rate, 1 aminocyclopropane-1-carboxylic acid oxidase (ACO) activity, and membrane permeability were assessed. After 7 months storage and 7 days shelf-life, apples subjected to 1-MCP and controlled atmosphere (CA) exhibited reduced ethylene production, respiration rate, and ACO activity compared to untreated or regular atmosphere counterparts. The combination of controlled atmosphere and pre-storage 1 MCP was the most effective in lowering ACO activity. Irrespective of conditions, 1 MCP curtailed ethylene production, respiration rate, and ACO activity during shelf-life, maintaining fruit firmness and slowing acidity loss. Only controlled atmosphere preserved quality and minimized disorders for optimally harvested apples, not late-harvested ones. None of the treatments maintained late-harvested apples quality after long-term storage plus shelf-life. The second experiment evaluated the effects of two ethanol vapor doses (250 or 500 ppm) or 1 MCP (650 ppb) with or without ethylene application (150 ppm) on the metabolism and quality of the apple cultivars ‘Elstar’ and ‘Nicoter’, over 14 d of holding at room temperature (20 ± 2 °C). For both cultivars studied, ethanol vapor treatments, especially 500 ppm, slowed the ripening of apples and inhibited the effect of applied ethylene on the ethylene production and respiration, but not as much as the 1 MCP treatment in ‘Nicoter’ apples. Ethanol application also resulted in higher succinate, malate and total organic acids concentrations. Ethanol application significantly reduced the sucrose conversion to glucose and fructose, while the ethanol + ethylene treatment resulted in high total sugars, fructose and sorbitol concentration after 14 d at 20 °C. The ethanol application (500 ppm) also affected conversion of succinate to fumarate, suggesting the succinate dehydrogenase activity as one possible action point of ethanol on the apple fruit metabolism. The combination of ethanol + ethylene treatments had a different response as compared to their isolated application, affecting sugars and organic acids metabolism differently. Fruit treated with ethanol vapor maintained lower electrolyte leakage, higher flesh firmness, greener color and had more sound fruit. However, its application increased the pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) activity and also enhanced acetaldehyde and ethyl acetate accumulation, but in concentrations below the odor threshold reported in the literature. The 1-MCP treatment increased decay incidence compared to the other treatments, reducing the amount of sound fruit in ‘Nicoter’ apples, but allowed higher acidity maintenance after 14 d holding at room temperature. There was no incidence of external and internal physiological disorders in either of the cultivars. In a third experiment, we explored the effects of the interaction between controlled atmosphere and 1-MCP treatment on ‘Alexander Lucas’ pears in storage, aiming to minimize internal storage disorders. Following treatment with 1-MCP at 300 ppb, the fruit were stored either at -0.5 or 1.0 °C in regular air or in CA (2.0 kPa O2 plus <0.7 kPa CO2). After six months of storage, superficial scald did not develop in fruit. The highest occurrence of flesh browning (72.2 %) was observed in air-stored fruit at -0.5 °C without 1-MCP treatment. Storage in regular air at 1.0 °C combined with 1-MCP resulted in 88.8 % sound fruit. Conversely, 1-MCP increased the incidence of flesh and core browning under CA conditions. Both 1-MCP and CA maintained greener skin color and higher titratable acidity. No significant differences were found for fruit firmness, total soluble solids and ascorbic acid content between the treatments. In conclusion, the quality of ‘Alexander Lucas’ pear was best maintained during six months storage under regular air at 1.0 °C combined with 300 ppb 1-MCP treatment.Publication Quantitative Proteomanalyse von Pseudomonaden zur Aufklärung biotechnologisch relevanter Stoffwechselwege(2013) Simon, Oliver; Huber, ArminThe main focus of this work was a quantitative proteome analysis of a variety of Pseudomonas strains with respect to the biotechnological synthesis of the base chemicals glyoxylic acid, butanol and vanillin. In addition, effects of the terpene citronellol on the proteome of P. aeruginosa were investigated. A second key aspect of this work involved the establishment of proteomics methods for the analysis of complex samples, especially for the analysis of membrane proteins. Using carbonate extraction followed by label-free MS-based quantification allowed the identification and quantification of a significant number of hydrophobic proteins which were not covered by the 2D-DIGE approach. In addition, the GeLCMSMS workflow was found to be a simple and efficient method for the analysis of total bacterial lysates. Using this method, about 30% of all proteins encoded by the P. putida KT2440 genome could be identified and quantified. In conclusion, this work demonstrated that different proteomics methods can substantially contribute to biotechnological strain development and the understanding of cellular networks.Publication Die Regulation der Futteraufnahme beim Schwein - Untersuchung der Wirkungen eines Serotonin Noradrenalin Wiederaufnahmehemmers (Sibutramin) und eines MCH-R1 Antagonisten (Compound B4)(2007) Sommer, Torsten; Claus, RolfThe regulation of food- and feed intake is a highly topical investigative area in animal husbandry as well as in human medicine. Here the focus of medicine is primarily on the reduction of food intake and at the same time change of the metabolism to increased energy expenditure in order to counteract the rising number of adiposity and metabolic syndrome. Therefore, substances from pharmacological research are known which act in different nuclei of the hypothalamus and control food intake and energy expenditure. In the present study two of these new substances were examined with regards to their impact on pigs. The results lead to a more detailed understanding of the corresponding mechanisms in pigs, one of the most important animals used in agriculture. In addition, the pig is also a suitable model for human pharmacological research, as it equals the human physiological regulation more than the rat, which is the most common animal model. Study work flow: Two different regulative substances were examined, on the one hand a serotonin noradrenalin reuptake inhibitor (Sibutramine) and on the other hand, a melanin concentrating hormone receptor antagonist (MCH-R1 antagonist; Compound B4). For the evaluation of the effectiveness the following factors were measured; feed intake, water consumption, weight gain, feed conversion rate and the development of body fat content. The changes caused in the regulation of the metabolism were evaluated by anabolic (Growth hormone and IGF-I) and catabolic (cortisol and aldosterone) hormones. In addition, thyroid hormones and urea in the blood plasma were measured as a parameter of protein turnover. The study was divided in three trials, each of them taking six weeks. During the first trial sibutramine was given per os to three animals and three animals were kept as control. During the second trial sibutramine was given intravenously to two animals and two animals were kept as control. During the third trial the MCH-R1 antagonist, Compound B4, was given per os to six animals. The results are described in the following sections. Results of the sibutramine application: The pharmacological effectiveness of sibutramine is mainly explained through its metabolites (desmethyl sibutramine and didesmethyl sibutramine). It was found that for the per os intake of 20 mg/kg LW sibutramine, as well as sibutramine as the first metabolite desmethyl-sibutramine, a lower concentration in the blood plasma existed compared to the intravenous application. Whereas concentration of the second metabolite didesmethyl-sibutramine was higher for per os application than for intravenous application. Since the effectiveness of sibutramine is mainly based on its second metabolite didesmethyl-sibutramine, the way of application was crucial for its effectiveness. This means that the intravenous application was ineffective. After per os application feed intake was significantly reduced, also dependent on the dose applied, but did not lead to a reduced weight gain of the animals. Further, no impact was found on the feed conversion ratio. In contrast, the treatment with sibutramine per os led to significantly increased water consumption. The application of sibutramine had no impact on the growth hormone secretion. However, IGF-1 concentration was increased while feed intake was decreased. The cortisol concentration in blood plasma was not impacted by any treatment. Results of the application of the MCH-R1 antagonist (Compound B4): The application of the antagonist led to a significant reduction of feed intake, dependent on the dose applied. The means derived for daily weight gain show considerable individual differences per animal, hence the reduction of daily weight gain overall was not significant. Further, changes in water consumption were also detected in this trial. However, it could not be clarified if this was caused by the application of the antagonist or the reduced feed intake. Generally, the application of Compound B4 tends to result in energy saving mechanisms, which have an impact on metabolic active hormones. IGF-1 for example, was slightly lower in the control phase. Neither sibutramine nor the MCH-R1 antagonist Compound B4 had any impact on the concentration of thyroid hormones or cortisol. Conclusions: The studies have shown that the MCH-R1 antagonist leads to a significant reduction of feed intake in pigs, whereas the metabolic active hormones, and therefore the energy expenditure, do not seem to be impacted at all. In addition, it was proven that the pig is a useful animal model for such human pharmacological investigations. However, the large amount of substances required can be seen as disadvantageous and as a limiting factor in the investigation of new agents with an initially small amount of substance synthesis.Publication Saponin fractions from fenugreek (Trigonella foenum-graecum L.)as dietary supplements for Nile tilapia (Oreochromis niloticus L.)and common carp (Cyprinus carpio L.)(2012) Stadtlander, Timo; Focken, UlfertThe future role and importance of aquaculture for the world food supply is more and more recognized by the public. High quality feed for semi-intensive and intensive aquaculture operations are necessary to guarantee a future continuous growth of fish production. Nutrient utilization efficiency has in the past been augmented by addition of feed additives, mainly anti-biotics and hormones. That practice is prohibited in the European Union since 1st of January 2006 and the consumer acceptance of products produced in that way is low. Around a decade ago saponins, secondary plant metabolites, have been found to be potential alternatives for aquafeeds. Saponinmixtures derived from the South American soap bark tree Quillaja saponaria, have been found to have beneficial effects on growth, feed and nutrient utilization and on metabolism of common carp Cyprinus carpio and Nile tilapia Oreochromis niloticus. Furthermore, in one of the early experiments a sex ratio in favor of males was observed in saponin fed tilapia compared to control fish not supplemented with saponins. An influence on sex ratio of Nile tilapia would be highly desirable since industrially Nile tilapia production is only financially viable if all male populations are produced since mixed sex tilapia populations tend to reproduce uncontrollable. The production of all male tilapia populations is mainly conducted by feeding the potentially environmentally hazardous and carcinogenic synthetic hormone 17--methyltestosterone to sexually undeveloped tilapia fry. The above mentioned problems led to a joint research project with partners in Israel and Palestine in which this dissertation has been conducted. The aim of the project was the fractionation of crude saponin extract derived from fenugreek Trigonella foenum-graecum and its testing in vitro and in vivo. It was assumed that a higher biological activity could be achieved if saponin fractions instead of crude mixtures would be applied. After in vitro testing, different saponin fractions or eluates, have been tested on their effects on growth, feed and nutrient utilization, metabolism, proximate composition, gene expression of GH and IGF-1, sex ratio, reproduction and gonad histology. Most of the feeding experiments were conducted with Nile tilapia but one experiment was also conducted with carp. Three of the feeding experiments have been conducted in a system capable of measuring continuous respiration of the fish. Another experiment has been conducted at a field station at Jericho, Palestine. The experiments conducted to evaluate the influence of saponin fractions on sex ratio of undifferentiated tilapia fry have been conducted primarily in a flow-through system. The tested saponin fractions and a tested sapogenin are not suitable to produce male monosex tilapia populations. The sex ratios after supplementation of diets of mixed sexed tilapia fry did only in one case show a significantly higher proportion of males than the control. In a larger scale repetition of that experiment the previous observations could statistically not be proofed. It must be assumed to be random effects or be the result of initially biased sex ratios after stocking undifferentiated tilapia fry. An experiment in which genetically female tilapias were fed with two fenugreek saponin fractions and positive and negative controls supported that finding. A long term feeding experiment conducted at Jericho revealed no influence of the long and short term supplemented Q. saponaria saponin mixtures on sex ratio and reproduction of genetically female tilapia. A similar laboratory experiment with fenugreek saponin fractions could not be evaluated in that regard. The experiments evaluating the effects of the saponin fractions on growth, feed and nutrient utilization, gene expression of GH and IGF-1 and proximate composition revealed no significant differences. But in all experiments one fraction eluated with 60%/40% (v/v) methanol/water showed numerically improved values compared to control and other fractions. Furthermore one fraction eluated with 40%/60%/ (v/v) methanol/water gave constantly numerically inferior results of tested parameters compared to control. That supports the conclusion that the 40% methanol fraction contains saponins generally referred to as anti-nutrients. In only one experiment, conducted at Jericho, significantly higher growth was observed after long term supplementation with a Q. saponaria saponin mixture containing elevated sapogenin content. The presented data in this thesis is not supporting an application of the tested saponin fractions as environmentally friendly alternative to methyltestosterone to produce all male populations of tilapia or to inhibit uncontrollable reproduction. Additional experiments are needed to evaluate different modes of application like immersion treatments or injections since during the experimental work of this thesis all tested saponins were added to the feed. An application of the tested fenugreek saponin fractions as growth promoters yields a higher potential although the experimental results are based upon low sample sizes due to capacity restrictions of the respirometric system. Therefore a repetition under near commercial or commercial conditions must be considered.