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Methanol based elongation of propionate and propionate and acetate in continuous and batch mixed culture systems
Smit, Sanne de; Leeuw, K.D. de; Buisman, C.J.N. ; Strik, D.P.B.T.B. - \ 2019
chain elongation - selective pressure - open-culture fermentation - mixed culture fermentation - biobased chemicals - methanol - butyrate - n-valerate
Continuous methanol based chain elongation of propionate to n-valerate was performed in a mixed culture reactor. Additionally, the study shows simultaneous methanol based elongation of propionate and acetate to respectively n-valerate and iso/n-butyrate in a continuous mixed culture reactor. A range of mixed culture batch experiments was performed with methanol and propionate and the initial pH ranging from 5 to 7.5 (steps of 0.5)
Importance of release location on the mode of action of butyrate derivatives in the avian gastrointestinal tract
Moquet, P.C.A. ; Onrust, L. ; Immerseel, F. Van; Ducatelle, R. ; Hendriks, W.H. ; Kwakkel, R.P. - \ 2016
Worlds Poultry Science Journal (2016). - ISSN 0043-9339 - p. 61 - 80.
broiler - butyrate - mode of action - release location
In the field of animal nutrition, butyrate is used as a zootechnical ingredient and can be used as an unprotected salt or in the form of protected derivatives such as butyrate glycerides or butyrate-loaded matrices. Dietary butyrate supplementation has been shown to improve growth performance and resilience of broiler chickens through distinct mechanisms, operating on both eukaryotic and prokaryotic cells. Firstly, butyrate influences endogenous avian cells in multiple ways: it is an agonist of free-fatty acid receptors, an inhibitor of pro-inflammatory pathways, an epigenetic modulating agent and acts as an energy source. Secondly, butyrate influences the microbiota residing in the avian gastrointestinal tract (GIT) as a result of its bacteriostatic properties. The responses, e.g. changes in growth performance, gut morphology, carcass traits or nutrient digestibility of chickens, to dietary butyrate supplementation are inconsistent with factors such as additive inclusion level, diet composition, age and health status of the bird modulating the effects of butyrate and its derivatives. For many derivatives, the precise GIT segment wherein butyrate is released is unclear. Release location may affect the observed responses to butyrate given the diversity of cell types and pH conditions encountered throughout the gastrointestinal tract of poultry, and the differences in microbiota composition in the different gut segments. As a consequence, our understanding of the mode of action of butyrate is hampered. Characterisation of existing derivatives and development of targeted-release formulations are, therefore, important to gain insight in the different physiological effects butyrate can elicit in broiler chickens.
Dietary Pectin-Derived Acidic Oligosaccharides Improve the Pulmonary Bacterial Clearance of Pseudomonas aeruginosa Lung Infection in Mice by Modulating Intestinal Microbiota and Immunity
Bernard, H. ; Desseyn, J.L. ; Bartke, N. ; Kleinjans, L.P.J. ; Belzer, C. ; Knol, J. ; Gottrand, F. ; Husson, M.O. - \ 2015
The Journal of Infectious Diseases 211 (2015)1. - ISSN 0022-1899 - p. 156 - 165.
cystic-fibrosis patients - chain fatty-acids - galacto-oligosaccharides - t-cells - cytokine production - virus-infection - human-milk - in-vitro - lactobacillus - butyrate
Background. A predominantly T-helper type 2 (Th2) immune response is critical in the prognosis of pulmonary Pseudomonas aeruginosa infection. But the mucosal and systemic immune responses can be influenced by the intestinal microbiota. Methods. We assessed the effect of microbiota compositional changes induced by a diet enriched in 5% acidic oligosaccharides derived from pectin (pAOS) on the immune response and outcome of chronic pulmonary P. aeruginosa infection in mice. Results. pAOS promoted Th1 polarization by increasing interferon ¿ release, upregulating t-bet gene expression, decreasing interleukin 4 secretion, and downregulating gata3 gene expression. pAOS also sustained the release of keratinocyte chemoattractant, recruited polynuclear leukocytes and macrophages, stimulated M1 macrophage activation and interleukin 10 release, and decreased tumor necrosis factor a release in the lung. These effects led to increased bacterial clearance after the first and second P. aeruginosa infections. pAOS modified the intestinal microbiota by stimulating the growth of species involved in immunity development, such as Bifidobacterium species, Sutturella wadsworthia, and Clostridium cluster XIVa organisms, and at the same time increased the production of butyrate and propionate. Conclusion. These results suggest that pAOS may have beneficial effects by limiting the number and severity of pulmonary exacerbations in patients chronically infected with P. aeruginosa, such as individuals with cystic fibrosis.
Aberrant gut microbiota composition at the onset of type 1 diabetes in young children
Goffau, M.C. de; Fuentes, S. ; Bogert, B. van den; Honkanen, H. ; Vos, W.M. de; Welling, G.W. ; Hyöty, H. ; Harmsen, H.J. - \ 2014
Diabetologia 57 (2014)8. - ISSN 0012-186X - p. 1569 - 1577.
intestinal microbiota - storage-conditions - disease - butyrate - age - community - immunity - health - bacteria - differs
Aims/hypothesis Recent studies indicate that an aberrant gut microbiota is associated with the development of type 1 diabetes, yet little is known about the microbiota in children who have diabetes at an early age. To this end, the microbiota of children aged 1–5 years with new-onset type 1 diabetes was compared with the microbiota of age-matched healthy controls. Methods A deep global analysis of the gut microbiota composition was established by phylogenetic microarray analysis using a Human Intestinal Tract Chip (HITChip). Results Principal component analyses highlighted the importance of age when comparing age-matched pairs. In pairs younger than 2.9 years, the combined abundance of the class Bacilli (notably streptococci) and the phylum Bacteroidetes was higher in diabetic children, whereas the combined abundance of members of Clostridium clusters IV and XIVa was higher in the healthy controls. Controls older than 2.9 years were characterised by a higher fraction of butyrate-producing species within Clostridium clusters IV and XIVa than was seen in the corresponding diabetic children or in children from the younger age groups, while the diabetic children older than 2.9 years could be differentiated by having an increased microbial diversity. Conclusions/interpretation The results from both age groups suggest that non-diabetic children have a more balanced microbiota in which butyrate-producing species appear to hold a pivotal position.
In Vitro fermentability of sugar beet pulp derived oligosaccharides using human and pig fecal inocula
Leijdekkers, A.G.M. ; Aguirre, M. ; Venema, K. ; Bosch, G. ; Gruppen, H. ; Schols, H.A. - \ 2014
Journal of Agricultural and Food Chemistry 62 (2014)5. - ISSN 0021-8561 - p. 1079 - 1087.
irritable-bowel-syndrome - pectic-oligosaccharides - arabino-oligosaccharides - ulcerative-colitis - large-intestine - fermentation - microbiota - butyrate - bacteria - profiles
The in vitro fermentation characteristics of different classes of sugar beet pectic oligosaccharides (SBPOS) were studied using human and pig fecal inocula. The SBPOS consisted mainly of partially acetylated rhamnogalacturonan-oligosaccharides and partially methyl esterified/acetylated homogalacturonan-oligosaccharides. Some SBPOS contained an unsaturated galacturonic acid residue at their non-reducing end. It was shown that SBPOS could be completely fermented by human and pig fecal microbiota, thereby producing butyrate, yet mainly acetate and propionate as metabolites. The degradation of SBPOS by pig fecal microbiota was different and much slower compared to human fecal microbiota. In general, rhamnogalacturonan-oligosaccharides were slower degraded than homogalacturonan32 oligosaccharides. Acetylation of rhamnogalacturonan-oligosaccharides lowered the degradation rate by pig fecal microbiota, but not by human fecal microbiota. No classic bifidogenic effect was shown for SBPOS using human fecal inoculum. However, several other potentially interesting modifications in the microbiota composition that can be associated with host health were observed, which are discussed.
Short-Chain Fatty Acids Activate AMP-Activated Protein Kinase and Ameliorate Ethanol-Induced Intestinal Barrier Dysfunction in Caco-2 Cell Monolayers
Eamin, E.E. ; Masclee, A.A. ; Dekker, J. ; Pieters, H.J. ; Jonkers, D.M. - \ 2013
The Journal of Nutrition 143 (2013)12. - ISSN 0022-3166 - p. 1872 - 1881.
alcoholic liver-disease - induced gut leakiness - paracellular permeability - signaling pathway - oxidative stress - epithelial-cells - colonic function - dietary fiber - leaky gut - butyrate
Short-chain fatty acids (SCFAs) have been shown to promote intestinal barrier function, but their protective effects against ethanol-induced intestinal injury and underlying mechanisms remain essentially unknown. The aim of the study was to analyze the influence of SCFAs on ethanol-induced barrier dysfunction and to examine the role of AMP-activated protein kinase (AMPK) as a possible mechanism using Caco-2 monolayers. The monolayers were treated apically with butyrate (2, 10, or 20 mmol/L), propionate (4, 20, or 40 mmol/L), or acetate (8, 40, or 80 mmol/L) for 1 h before ethanol (40 mmol/L) for 3 h. Barrier function was analyzed by measurement of transepithelial resistance and permeation of fluorescein isothiocyanate-labeled dextran. Distribution of the tight junction (TJ) proteins zona occludens-1, occludin, and filamentous-actin (F-actin) was examined by immunofluorescence. Metabolic stress was determined by measuring oxidative stress, mitochondrial function, and ATP using dichlorofluorescein diacetate, dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide, and bioluminescence assay, respectively. AMPK was knocked down by small interfering RNA (siRNA), and its activity was assessed by a cell-based ELISA. Exposure to ethanol significantly impaired barrier function compared with controls (P <0.0001), disrupted TJ and F-actin cytoskeleton integrity, and induced metabolic stress. However, pretreatment with 2 mmol/L butyrate, 4 mmol/L propionate, and 8 mmol/L acetate significantly alleviated the ethanol-induced barrier dysfunction, TJ and F-actin disruption, and metabolic stress compared with ethanol-exposed monolayers (P <0.0001). The promoting effects on barrier function were abolished by inhibiting AMPK using either compound C or siRNA. These observations indicate that SCFAs exhibit protective effects against ethanol-induced barrier disruption via AMPK activation, suggesting a potential for SCFAs as prophylactic and/or therapeutic factors against ethanol-induced gut leakiness.
Selective carboxylate production by controlling hydrogen, carbon dioxide and substrate concentrations in mixed culture fermentation
Arslan, D. ; Steinbusch, K.J.J. ; Diels, L. ; Wever, H. de; Hamelers, H.V.M. ; Buisman, C.J.N. - \ 2013
Bioresource Technology 136 (2013). - ISSN 0960-8524 - p. 452 - 460.
sequencing batch reactor - biohydrogen production - biological production - waste-water - bacteria - glucose - acid - bioconversion - butyrate - caproate
This research demonstrated the selective production of n-butyrate from mixed culture by applying 2 bar carbon dioxide into the headspace of batch fermenters or by increasing the initial substrate concentration. The effect of increasing initial substrate concentration was investigated at 8, 13.5 and 23 g COD/L with potato processing waste stream. Within 1 week of incubation, n-butyrate fraction selectively increased up to 83% by applying 2 bar hydrogen or 78% by applying carbon dioxide into the headspace whereas it was only 59% in the control reactor. Although the fraction of n-butyrate was elevated, the concentration remained lower than in the control. Both the highest concentration and fraction of n-butyrate were observed under the highest initial substrate concentration without headspace addition. The concentration was 10 g COD/L with 73% fraction. The operational conditions obtained from batch experiments for selective n-butyrate production were validated in a continuous process.
Short chain fatty acids stimulate Angiopoietin-like 4 synthesis in human colon adenocarcinoma cells by activating PPARy
Alex, S. ; Lange, K. ; Amolo, T. ; Grinstead, J.S. ; Haakonsson, A.K. ; Szalowska, E. ; Koppen, A. ; Mudde, C.M. ; Haenen, D. ; Al-Lahham, S. ; Roelofsen, H. ; Houtman, R. ; Burg, B. van der; Mandrup, S. ; Bonvin, A.M.J.J. ; Kalkhoven, E. ; Muller, M.R. ; Hooiveld, G.J.E.J. ; Kersten, A.H. - \ 2013
Molecular and Cellular Biology 33 (2013)7. - ISSN 0270-7306 - p. 1303 - 1316.
inflammatory-bowel-disease - ppar-gamma - transcriptional activity - lipoprotein-lipase - skeletal-muscle - gut microbiota - target gene - expression - protein-4 - butyrate
Angiopoietin-like protein 4 (ANGPTL4/FIAF) has been proposed as a circulating mediator between the gut microbiota and fat storage. Here, we show that transcription and secretion of ANGPTL4 in human T84 and HT29 colon adenocarcinoma cells is highly induced by physiological concentrations of short-chain fatty acids (SCFA). SCFA induce ANGPTL4 by activating the nuclear receptor peroxisome proliferator activated receptor ¿ (PPAR¿), as demonstrated using PPAR¿ antagonist, PPAR¿ knockdown, and transactivation assays, which show activation of PPAR¿ but not PPARa and PPARd by SCFA. At concentrations required for PPAR¿ activation and ANGPTL4 induction in colon adenocarcinoma cells, SCFA do not stimulate PPAR¿ in mouse 3T3-L1 and human SGBS adipocytes, suggesting that SCFA act as selective PPAR¿ modulators (SPPARM), which is supported by coactivator peptide recruitment assay and structural modeling. Consistent with the notion that fermentation leads to PPAR activation in vivo, feeding mice a diet rich in inulin induced PPAR target genes and pathways in the colon. We conclude that (i) SCFA potently stimulate ANGPTL4 synthesis in human colon adenocarcinoma cells and (ii) SCFA transactivate and bind to PPAR¿. Our data point to activation of PPARs as a novel mechanism of gene regulation by SCFA in the colon, in addition to other mechanisms of action of SCFA.
Bioelectrochemical production of caproate and caprylate from acetate by mixed cultures
Eerten-Jansen, M.C.A.A. van; Heijne, A. ter; Grootscholten, T.I.M. ; Steinbusch, K.J.J. ; Sleutels, T.H.J.A. ; Hamelers, H.V.M. ; Buisman, C.J.N. - \ 2013
ACS sustainable chemistry & engineering 1 (2013)5. - ISSN 2168-0485 - p. 513 - 518.
microbial electrolysis cells - fuel-cells - hydrogen - biomass - conversion - ethanol - reduction - transport - membranes - butyrate
The use of mixed cultures to convert waste biomass into medium chain fatty acids, precursors for renewable fuels or chemicals, is a promising route. To convert waste biomass into medium chain fatty acids, an external electron donor in the form of hydrogen or ethanol needs to be added. This study investigated whether the cathode of a bioelectrochemical system can be used as the electron donor for the conversion of acetate into medium chain fatty acids. We show that medium chain fatty acids were produced in a bioelectrochemical system at -0.9 V vs. NHE cathode potential, without addition of an external mediator. Caproate, butyrate and smaller fractions of caprylate were the main products formed from acetate. In-situ produced hydrogen was likely involved as an electron donor for the reduction of acetate. Electron and carbon balances revealed that 45% of the electrons in electric current and acetate, and 31% of the carbon from acetate were recovered in the formed products. This study showed for the first time production of medium chain fatty acids caproate and caprylate from acetate at the cathode of bioelectrochemical systems, and offers new opportunities for application of bioelectrochemical systems.
Iron Depletion and Repletion with Ferrous Sulfate or Electrolytic Iron Modifies the Composition and Metabolic Activity of the Gut Microbiota in Rats
Dostal, A. ; Chassard, C. ; Hilty, F.M. ; Zimmermann, M.B. ; Jaeggi, T. ; Rossi, S. ; Lacroix, C. - \ 2012
The Journal of Nutrition 142 (2012)2. - ISSN 0022-3166 - p. 271 - 277.
bacteroides-fragilis - human colon - butyrate - bacteria - health - flora - requirement - growth - conservation - lactobacilli
Iron (Fe) deficiency anemia is a global health concern and Fe fortification and supplementation are common corrective strategies. Fe is essential not only for the human host but also for nearly all gut bacteria. We studied the impact of Fe deficiency and Fe repletion on the gut microbiota in rats. Weanling rats were fed an Fe-deficient diet for 24 d and then repleted for 13 d with FeSO(4) (n = 15) or electrolytic Fe (n = 14) at 10 and 20 mg Fe . kg diet(-1). In addition, one group of rats (n = 8) received the Fe-deficient diet and one group In = 3) received a Fe-sufficient control diet for all 37 d. Fecal samples were collected at baseline and after the depletion and repletion periods, and colonic tissues were examined histologically. Microbial metabolite composition in cecal water was measured and fecal samples were analyzed for microbial composition with temporal temperature gradient gel electrophoresis and qPCR. Compared to Fe-sufficient rats, Fe-deficient rats had significantly lower concentrations of cecal butyrate (-87%) and propionate (-72%) and the abundance of dominant species was strongly modified, including greater numbers of lactobacilli and Enterobacteriaceae and a large significant decrease of the Roseburia spp./E. rectale group, a major butyrate producer. Repletion with 20 mg FeSO(4) . kg diet(-1) significantly increased cecal butyrate concentrations and partially restored bacterial populations compared to Fe-deficient rats at endpoint. The effects on the gut microbiota were stronger in rats repleted with FeSO(4) than in rats repleted with electrolytic Fe, suggesting ferrous Fe may be more available for utilization by the gut microbiota than elemental Fe. Repletion with FeSO(4) significantly increased neutrophilic infiltration of the colonic mucosa compared to Fe-deficient rats. In conclusion, Fe depletion and repletion strongly affect the composition and metabolic activity of rat gut microbiota. J. Nutr. 142: 271-277, 2012,
Transfer of Intestinal Microbiota From Lean Donors Increases Insulin Sensitivity in Individuals With Metabolic Syndrome
Vrieze, A. ; Nood, E. van; Holleman, F. ; Heilig, G.H.J. ; Zoetendal, E.G. ; Vos, W.M. de - \ 2012
Gastroenterology 143 (2012)4. - ISSN 0016-5085 - p. 913 - 916.
gut microbiome - butyrate - obesity - resistance - bacteria - storage - tract - probe
Alterations in intestinal microbiota are associated with obesity and insulin resistance. We studied the effects of infusing intestinal microbiota from lean donors to male recipients with metabolic syndrome on the recipients' microbiota composition and glucose metabolism. Subjects were assigned randomly to groups that were given small intestinal infusions of allogenic or autologous microbiota. Six weeks after infusion of microbiota from lean donors, insulin sensitivity of recipients increased (median rate of glucose disappearance changed from 26.2 to 45.3 µmol/kg/min; P <.05) along with levels of butyrate-producing intestinal microbiota. Intestinal microbiota might be developed as therapeutic agents to increase insulin sensitivity in humans; www.trialregister.nl; registered at the Dutch Trial Register (NTR1776)
Profiling human gut bacterial metabolism and its kinetic using [U-(13)C]glucose and NMR
Graaf, A.A. de; Maathuis, A. ; Waard, P. de; Deutz, N.E.P. ; Dijkema, C. ; Vos, W.M. de; Venema, K. - \ 2010
NMR in Biomedicine 23 (2010)1. - ISSN 0952-3480 - p. 2 - 12.
chain fatty-acids - amino-acids - fermentation products - large-intestine - colon-cancer - human feces - butyrate - ethanol - spectrometry - acetaldehyde
This study introduces a stable-isotope metabolic approach employing [U-(13)C]glucose that, as a novelty, allows selective profiling of the human intestinal microbial metabolic products of carbohydrate food components, as well as the measurement of the kinetics of their formation pathways, in a single experiment. A well-established, validated in vitro model of human intestinal fermentation was inoculated with standardized gastrointestinal microbiota from volunteers. After culture stabilization, [U-(13)C]glucose was added as an isotopically labeled metabolic precursor. System lumen and dialysate samples were taken at regular intervals. Metabolite concentrations and isotopic labeling were determined by NMR, GC, and enzymatic methods. The main microbial metabolites were lactate, acetate, butyrate, formate, ethanol, and glycerol. They together accounted for a (13)C recovery rate as high as 91.2%. Using an NMR chemical shift prediction approach, several minor products that showed (13)C incorporation were identified as organic acids, amino acids, and various alcohols. Using computer modeling of the (12)C contents and (13)C labeling kinetics, the metabolic fluxes in the gut microbial pathways for synthesis of lactate, formate, acetate, and butyrate were determined separately for glucose and unlabeled background substrates. This novel approach enables the study of the modulation of human intestinal function by single nutrients, providing a new rational basis for achieving control of the short-chain fatty acids profile by manipulating substrate and microbiota composition in a purposeful manner.
Parenteral long-acting amoxicillin reduces intestinal bacterial community diversity in piglets even 5 weeks after the administration
Janzcyk, P. ; Pieper, R. ; Souffrant, W.B. ; Bimczok, D. ; Rothkotter, H.J. ; Smidt, H. - \ 2007
ISME Journal 1 (2007). - ISSN 1751-7362 - p. 180 - 183.
gradient gel-electrophoresis - weaning piglets - human feces - sp nov. - microbiota - antibiotics - butyrate - abundant - dietary - pigs
We investigated the long-term effects of a single intramuscular administration of amoxicillin (15 mg kg-1) 1 day after birth, on piglet intestinal microbiota. Animals received no creep feed before weaning on day 28 of age. For the next 11 days, the piglets received a wheat¿barley-based diet. Colon digesta samples were collected on day 39 and subjected to denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene fragments. DGGE fingerprint diversity indices differed between the group treated with amoxicillin and the untreated group (0.80.19 and 1.030.17, respectively, P=0.012). Reamplification and sequencing of two bands present in all samples revealed that a Roseburia faecalis-related population was strongly reduced in relative abundance (98% identity) in the treated group, while an enterobacterial population with 100% identity to Shigella spp., Escherichia coli and Salmonella enterica serovar Typhi was enriched. A band corresponding to Lactobacillus sobrius was present only in the control group. The protective effect of prophylactic antibiotic administration may be outweighed by the long-lasting disturbance of the gut ecosystem.