Browsing by Subject "Metabolites"

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Characterization of host and bacterial proteins in crossbred grower pigs at marginal lysine concentration
(2025) Kurz, Alina; Seifert, Jana
The current pork production strategies still result in high nitrogen emissions and thus harmful effects on the environment. The animals excrete about two-thirds of the protein in the diet unused, which increases nitrogen emissions and makes feeding the animals according to their needs expensive and inefficient. To minimize the negative aspects of pork production, it is important to reduce the use of proteins in feed while improving the protein efficiency of pigs. Improving protein efficiency requires a comprehensive understanding of physiological processes, particularly enzymatic activities, amino acid transporter capacities, the proteome, the metabolome and the microbiome. The aim of this work was therefore to identify animal- specific and quantitatively expressed proteins, enzymes, amino acid transporters and metabolites and to correlate these findings with protein turnover data. The study also examined whether the efficiency of protein utilization is influenced by these variables. A total of 48 male crossbred animals (German Landrace x Pietrain) were slaughtered in the 21st week of life for the present experiment. The animals were kept individually under the same conditions from the 75th day of life until the day of slaughter. A two-phase ad libitum feeding was carried out, with the change from phase 1 to phase 2 taking place in the 14th week of life. Since the genetic potential of protein utilization efficiency was to be estimated, the content of the first limiting amino acid lysine was 90 %, just below the GfE (2006) requirement limit in order to ensure a limiting factor for the protein approach. Thus, lysine was the limiting factor for protein retention and allowed animals to exploit their full genetic potential for protein utilization. After euthanizing, the animals were opened, and the stomach, small intestine and large intestine removed. Mucosal samples were taken from the sections of the pars nonglandularis, cardiac gland zone and pars pylorica, as well as digesta samples from the stomach. In addition, digesta and mucosal samples were collected from the duodenum, jejunum, ileum, colon, and cecum. The proteome in all samples was analyzed. The metabolome and enzyme activities were determined in all digesta samples. In addition, amino acid transporter expression was examined in all small intestinal mucosal samples. The activity of the enzymes trypsin, chymotrypsin, carboxypeptidase A and carboxypeptidase B were examined in the digesta samples from all sections. The highest activities were found in the small intestine, the lowest activities in the stomach and large intestine. The trypsin and chymotrypsin activity was highest in the jejunum, whereas the activities of the carboxypeptidases were highest in the duodenum. In addition, individual animal differences in enzyme activities could be identified. In addition, the expression rate of various amino acid transporters was examined in the present work. In the duodenum the transporter SLC1A5 showed the highest expression rate, in the jejunum the transporters SLC5A1 and SLC6A19. In the ileum, the rates of the transporters SLC1A1, SLC7A1 and SLC7A9 were at the highest level. The identification of the host proteome revealed different protein patterns in the examined sections of the gastrointestinal tract. In particular, host proteins were identified in the mucosa samples, with the exception of those from the ileum and cecum. The high number of host proteins in the mucosa samples, which are associated with cellular processes and metabolism, underlines its role in physiological digestive processes. In addition, the examined host proteins could be assigned to numerous KEGG pathways, thus creating a deeper understanding of physiological and metabolic pathways. The study of the bacterial proteome allowed not only categorization into different metabolic pathways, but also identification of the active microbiome in the different sections. For example, it was shown that Firmicutes dominate in the stomach and small intestine, whereas Bacteroidetes are mainly found in the large intestine. This highlights the different roles of the sections examined, with Firmicutes being primarily responsible for breaking down proteins and carbohydrates and Bacteroidetes playing a central role in the fermentation of undigested proteins and carbohydrates in the large intestine. Additionally, the increased presence of Tenericutes in the mucosal samples indicated a possible specialization of this family to the conditions in the mucosal environment, where they interact with host cells and contribute to metabolic processes. The identification of numerous unclassified bacterial groups also shows how complex the composition of the microbiome is and that further research must be carried out in order to be able to fully identify connections. The examination of the various metabolites in the digesta samples from the gastrointestinal tract showed clear differences in the detection rate. While the fatty acids acetate, butyrate, valerate, isobutyrate and propionate were found primarily in the sections of the large intestine, lactate, isovalerate and the amino acids showed a higher occurrence in the sections of the small intestine. This is in line with the scientific knowledge that fatty acids are primarily formed during microbial fermentation in the large intestine and should therefore occur in higher quantities here. Overall, the results of the present work showed that, in addition to physiological differences between the sections of the gastrointestinal tract, there also appear to be significant animal- specific differences in all parameters examined. These differences can have an influence on the efficiency of the animals, but it is still important to find out which factors cause these differences. Since the feeding, husbandry concept and experimental design were chosen in such a way that there is as little variation and discrepancy between the animals as possible, a genetic influence on the efficiency of the animals cannot be ruled out. If the animals' genetic potential to use nitrogen efficiently becomes apparent, a further foundation stone has been laid for making pork production tailored to needs and conserving resources and the environment.
Multi-omics reveals different strategies in the immune and metabolic systems of high-yielding strains of laying hens
(2022) Iqbal, Muhammad Arsalan; Reyer, Henry; Oster, Michael; Hadlich, Frieder; Trakooljul, Nares; Perdomo-Sabogal, Alvaro; Schmucker, Sonja; Stefanski, Volker; Roth, Christoph; Camarinha-Silva, Amélia; Huber, Korinna; Sommerfeld, Vera; Rodehutscord, Markus; Wimmers, Klaus; Ponsuksili, Siriluck
Lohmann Brown (LB) and Lohmann Selected Leghorn (LSL) are two commercially important laying hen strains due to their high egg production and excellent commercial suitability. The present study integrated multiple data sets along the genotype-phenotype map to better understand how the genetic background of the two strains influences their molecular pathways. In total, 71 individuals were analyzed (LB, n = 36; LSL, n = 35). Data sets include gut miRNA and mRNA transcriptome data, microbiota composition, immune cells, inositol phosphate metabolites, minerals, and hormones from different organs of the two hen strains. All complex data sets were pre-processed, normalized, and compatible with the mixOmics platform. The most discriminant features between two laying strains included 20 miRNAs, 20 mRNAs, 16 immune cells, 10 microbes, 11 phenotypic traits, and 16 metabolites. The expression of specific miRNAs and the abundance of immune cell types were related to the enrichment of immune pathways in the LSL strain. In contrast, more microbial taxa specific to the LB strain were identified, and the abundance of certain microbes strongly correlated with host gut transcripts enriched in immunological and metabolic pathways. Our findings indicate that both strains employ distinct inherent strategies to acquire and maintain their immune and metabolic systems under high-performance conditions. In addition, the study provides a new perspective on a view of the functional biodiversity that emerges during strain selection and contributes to the understanding of the role of host–gut interaction, including immune phenotype, microbiota, gut transcriptome, and metabolome.
Regulatory modules of metabolites and protein phosphorylation in arabidopsis genotypes with altered sucrose allocation
(2022) Stefan, Thorsten; Wu, Xu Na; Zhang, Youjun; Fernie, Alisdair; Schulze, Waltraud X.
Multi-omics data sets are increasingly being used for the interpretation of cellular processes in response to environmental cues. Especially, the posttranslational modification of proteins by phosphorylation is an important regulatory process affecting protein activity and/or localization, which, in turn, can have effects on metabolic processes and metabolite levels. Despite this importance, relationships between protein phosphorylation status and metabolite abundance remain largely underexplored. Here, we used a phosphoproteomics–metabolomics data set collected at the end of day and night in shoots and roots of Arabidopsis to propose regulatory relationships between protein phosphorylation and accumulation or allocation of metabolites. For this purpose, we introduced a novel, robust co-expression measure suited to the structure of our data sets, and we used this measure to construct metabolite-phosphopeptide networks. These networks were compared between wild type and plants with perturbations in key processes of sugar metabolism, namely, sucrose export (sweet11/12 mutant) and starch synthesis (pgm mutant). The phosphopeptide–metabolite network turned out to be highly sensitive to perturbations in sugar metabolism. Specifically, KING1, the regulatory subunit of SnRK1, was identified as a primary candidate connecting protein phosphorylation status with metabolism. We additionally identified strong changes in the fatty acid network of the sweet11/12 mutant, potentially resulting from a combination of fatty acid signaling and metabolic overflow reactions in response to high internal sucrose concentrations. Our results further suggest novel protein-metabolite relationships as candidates for future targeted research.
Spent Pleurotus ostreatus substrate has potential for managing Fusarium wilt of banana
(2021) Ocimati, Walter; Were, Evans; Tazuba, Anthony Fredrick; Dita, Miguel; Zheng, Si-Jun; Blomme, Guy
A range of basidiomycetes including the edible mushroom Pleurotus ostreatus (Po) can suppress plant pathogens such as Fusarium spp. With the current increase in production and consumption of Po in Uganda, the spent Po substrate (SPoS) could be an alternative to manage Fusarium wilt of banana (FWB), caused by the soil borne pathogen Fusarium oxysporum f. sp. cubense, race 1 (Foc). This study determined the potential of SPoS to inhibit Foc in vitro and in potted plants. In vitro studies confirmed suppression of Foc in pure co-culture (Po vs. Foc) assays and media amended with different concentrations (0% to 50% w/v) of un-sterilized SPoS filtrates. Foc growth in the sterile SPoS filtrate was comparable to the water control, suggesting possible roles of biotic or thermolabile components of the SPoS. To further verify the suppressive effects of SPoS, pot experiments were carried out with a resistant (‘Mbwazirume’, AAA) and susceptible (‘Sukali Ndizi’, AAB) banana cultivar using both artificially and naturally infested soils. Independent of the inoculation method, SPoS significantly reduced the severity of FWB in pot experiments. Susceptible cultivar ‘Sukali Ndizi’ growing in substrates amended with SPoS showed lower (1.25) corm damage (Scale 0–5) than the un-amended control (3.75). No corm damage was observed in uninoculated controls. The resistant cultivar ‘Mbwazirume’, showed slight (0.25) corm damage only in the Foc-inoculated plants without SPoS. These findings suggest that SPoS could be used as part of the management practices to reduce the impact of FWB.
Synthesis of human phase I and phase II metabolites of hop (Humulus lupulus) prenylated flavonoids
(2022) Buckett, Lance; Schönberger, Sabrina; Spindler, Veronika; Sus, Nadine; Schoergenhofer, Christian; Frank, Jan; Frank, Oliver; Rychlik, Michael
Hop prenylated flavonoids have been investigated for their in vivo activities due to their broad spectrum of positive health effects. Previous studies on the metabolism of xanthohumol using untargeted methods have found that it is first degraded into 8-prenylnaringenin and 6-prenylnaringenin, by spontaneous cyclisation into isoxanthohumol, and subsequently demethylated by gut bacteria. Further combinations of metabolism by hydroxylation, sulfation, and glucuronidation result in an unknown number of isomers. Most investigations involving the analysis of prenylated flavonoids used surrogate or untargeted approaches in metabolite identification, which is prone to errors in absolute identification. Here, we present a synthetic approach to obtaining reference standards for the identification of human xanthohumol metabolites. The synthesised metabolites were subsequently analysed by qTOF LC-MS/MS, and some were matched to a human blood sample obtained after the consumption of 43 mg of micellarised xanthohumol. Additionally, isomers of the reference standards were identified due to their having the same mass fragmentation pattern and different retention times. Overall, the methods unequivocally identified the metabolites of xanthohumol that are present in the blood circulatory system. Lastly, in vitro bioactive testing should be applied using metabolites and not original compounds, as free compounds are scarcely found in human blood.