Composition Of Short-chain Fatty Acids Research Articles

Dietary Complex Probiotic Supplementation Changed the Composition of Intestinal Short-Chain Fatty Acids and Improved the Average Daily Gain of Growing Pigs

At present, probiotics are being extensively evaluated for their efficacy as an alternative to antibiotics, and their safety in livestock production. In this study, 128 (Duroc, Yorkshire and Landrace) pigs with an average initial body weight of 28.38 ± 0.25 kg were allocated to four dietary treatments in a randomized complete-block design. There were eight pens per treatment, with four pigs per pen (two barrows and two gilts). Dietary treatments included: (1) control diet; (2) control diet + 0.05% complex probiotic; (3) control diet + 0.1% complex probiotic; (4) control diet + 0.2% complex probiotic. During the 28-day experimental period, the feeding of 0.1% complex probiotic in the diet increased body weight and average daily gain (p < 0.05). The addition of complex probiotics decreased total cholesterol and glucose concentrations in the blood (p < 0.01). Acetate concentrations in the blood increased from 0.1% complex probiotic in the diet (p < 0.05), while NH3 and H2S emissions in the feces decreased (p < 0.05) from 0.1% or 0.2% complex probiotic in the diet. In conclusion, dietary complex probiotic supplementation changed the composition of intestinal short-chain fatty acids and improved growth performance for growing pigs.

Advances in prebiotics for poultry: role of the caeca and oligosaccharides

Prebiotics are non-digestible carbohydrates that selectively stimulate the growth of beneficial bacteria. Prebiotic supplementation into poultry diets results in a decreased rate of pathogenic bacteria colonisation in the gastrointestinal tract. It also enhances production of volatile fatty acids and lactic acid, which provide the bird with energy. This results in improved host gastrointestinal health and productive performance. Oligosaccharides are the most notable prebiotics in poultry nutrition. Examples of prebiotic oligosaccharides include xylo-oligosaccharides, fructo-oligosaccharides, and galacto-oligosaccharides. Oligosaccharides are derived from hydrolysis of non-starch polysaccharides (NSP). They are manufactured from plant sources, synthesised by physiochemical methods or enzymatic processes. The effects of oligosaccharides occur primarily in the caeca; oligosaccharides bypass the small intestine and reach the caeca, where they are readily fermented by beneficial bacteria, such as those in family Lactobacillaceae and Bifidobacteriaceae. Caeca function is generally poorly understood, despite extensive reviews and studies in this field. A deeper understanding of the factors that influence ability of the caeca to effectively utilise oligosaccharides is warranted. This would allow new prebiotic products and NSP- degrading enzymes to be developed, targeted to specific diets and scenarios. This is required, given the lack of consistency observed in the outputs derived from different studies assessing oligosaccharide efficacy in poultry diets. A key hinderance to progression in this field is that authors rarely analyse the oligosaccharide content and composition in the test diets and products, or in the bird’s gastrointestinal tract. This review examines the mechanisms behind how oligosaccharides induce prebiotic effects in poultry, by identifying the role of the caeca in NSP digestion and identifying the impact of oligosaccharides on caeca microbiota and short-chain fatty acid composition.

Fecal microbiota transplantation and replenishment of short-chain fatty acids protect against chronic cerebral hypoperfusion-induced colonic dysfunction by regulating gut microbiota, differentiation of Th17 cells, and mitochondrial energy metabolism

BackgroundLittle is known about the association between gut microbiota and intestinal injury under a state of chronic cerebral hypoperfusion (CCH). Here, the effects of gut microbiota and short-chain fatty acids (SCFAs), as important metabolic products, on intestinal function and potential mechanisms after CCH were investigated.MethodsRats were subjected to bilateral common carotid artery occlusion (BCCAo) to induce CCH. The gut microbiota and metabolites of SCFAs were assessed by 16S rRNA sequencing and targeted metabolomics, respectively. Transcriptomic analysis of colon tissues was also conducted. Subsequently, potential molecular pathways and differentially expressed genes were verified by western blot, immunoprecipitation, and immunofluorescence analyses. Furthermore, the integrity of the colonic barrier was evaluated by hematoxylin and eosin and mucin 2 staining and expression levels of tight junction proteins. Besides, colonic inflammation was further assessed by flow cytometry and expression levels of inflammatory cytokines. In addition, colonic mitochondrial dysfunction was analyzed via membrane potential, reactive oxygen species, electron transport chain (ETC) activities, adenosine triphosphate content, and mitochondrial ultrastructure.ResultsCCH modified gut microbial composition and microbial metabolism of SCFAs, which may be associated with inhibition of mitochondrial ETC activities and oxidative phosphorylation, leading to dysregulation of mitochondrial energy metabolism. Furthermore, CCH induced differentiation of pathogenic Th17 cells, promoted the formation of complexes of interferon regulatory factor 4 and signal transducer and activator of transcription 3 (STAT3), and increased the phosphorylation of STAT3. This was associated with an impairment of colonic barrier function and chronic colonic inflammation. In contrast, FMT and SCFA replenishment ameliorated CCH-induced gut microbial dysbiosis by increasing the intestinal content of Ruminococcus_sp_N15_MGS_57 and modulating microbial metabolism of SCFAs by increasing acetic acid contents associated with an improvment of the balance between Tregs and Th17 cells, mitochondrial ETC activities, and oxidative phosphorylation to prevent colonic inflammation and dysregulation of mitochondrial energy metabolism.ConclusionThese findings indicate that FMT and SCFA replenishment present a promising therapeutic strategy against colonic dysfunction under a state of chronic cerebral ischemia.

Alterations of the gut microbiota and short chain fatty acids in necrotizing enterocolitis and food protein-induced allergic protocolitis infants: A prospective cohort study.

Even though presenting with similar clinical manifestations, necrotizing enterocolitis (NEC) and food protein-induced allergic protocolitis (FPIAP) have completely different treatments and prognosis. Our study aimed to quantify and evaluate differences in gut microbiota and short chain fatty acids (SCFAs) between infants with NEC and FPIAP to better identify these two diseases in clinical settings. A total of 43 infants with NEC or FPIAP in Children's Hospital of Chongqing Medical University, China between December 2020 and December 2021 were enrolled. Stool samples were prospectively collected and froze. Infants defined as NEC were those who presented with clinical courses consistent with NEC and whose radiographs fulfilled criteria for Bell's stage 2 or 3 NEC, while those who were healthy in appearance and had blood in the stool (visible or may be microscopic), had normal bowel sounds in physical examination, were resolved after eliminating the causative food, and/or had recurrence of symptoms after oral food challenge (OFC) were defined as FPIAP. Primers specific for bacterial 16S rRNA genes were used to amplify and pyrosequence fecal DNA from stool samples. Gas chromatography-mass spectrometry (GC-MS) technology was used to determine the concentrations of SCFAs. Among the 43 infants, 22 were diagnosed with NEC and 21 were diagnosed with FPIAP. The microbial community structure in NEC infant stools differed significantly from those in FPIAP infant stools. NEC infants had significantly higher proportion of Actinobacteria and reduced proportion of Bacteroidetes compared with FPIAP infants, and the proportions of Halomonas, Acinetobacter, Bifidobacterium, and Stenotrophomonas in NEC infants were significantly higher than that of FPIAP infants. In addition, infants with NEC had significantly lower levels of acetic acid, propionic acid, butyric acid, isovaleric acid, and total SCFAs, and higher level of hexanoic acid as compared to the infants of the FPIAP group. The differences of gut microbiota composition and concentrations of SCFAs might represent suitable biomarker targets for early identification of NEC and FPIAP.

Effects on feed intake, milk production, and methane emission in dairy cows fed silage or fresh grass with concentrate or fresh grass harvested at early or late maturity stage without concentrate.

The objective of the study was to quantify the effects on dry matter intake (DMI), nutrient digestibility, gas exchange, milk production, and milk quality in dairy cows fed fresh grass harvested at different maturity stages. Sixteen Danish Holstein cows in mid-lactation were divided into 4 blocks and used in 4 incomplete 4 × 2 Latin squares with 2 periods of 21 d. The cows received 1 of 4 treatments in each period, resulting in 8 cows per treatment, as follows: grass-clover silage supplemented with 6 kg/d concentrate pellets (SILc), fresh grass harvested at late maturity stage supplemented with 6 kg/d concentrate pellets (LATc), fresh grass harvested at late maturity stage (LAT), and fresh grass harvested at early maturity stage (ERL). The cows were housed in tiestalls and milked twice daily. The cows had ad libitum access to the forage, and concentrate pellets were divided into equal amounts and fed separately in the morning and afternoon. Fecal samples were collected to determine apparent total-tract digestibility, and samples of rumen fluid were collected for determination of short chain fatty acid composition. Halters were used for measuring eating and rumination time. Gas exchange was measured in open-circuit respiration chambers. Total DMI was higher in LATc and ERL (16.9 ± 0.45 and 15.5 ± 0.39 kg/d, respectively) compared with LAT (14.1 ± 0.42 kg/d). Relative to SILc, cows fed fresh grass experienced a convex pattern in DMI during the experiment. The changes in DMI were related to changes in leaf to stem ratio, fiber concentration, and organic matter digestibility determined in vitro in samples of the fresh grass harvested throughout the experiment. The apparent total-tract digestibility of organic matter was higher in SILc and LAT compared with LATc. Methane yield was lower for LATc compared with LAT (19.5 ± 0.61 vs. 22.6 ± 0.55 g of CH4/kg of DMI), and was not different between LAT and ERL. Compared with LAT, milk yield was higher for ERL (21.1 ± 1.14 vs. 23.4 ± 1.11 kg/d) and energy-corrected milk (ECM) yield was higher for LATc (21.5 ± 0.99 vs. 25.3 ± 1.03 kg/d). We detected no differences in milk or ECM yield between SILc and LATc. Milk protein yield was higher and milk fat concentration was lower in LATc compared with LAT. The fatty acid percentages of ∑C4-C14:1 and ∑C16 in milk were higher for SILc compared with LATc, signifying pronounced de novo synthesis. The n-6:n-3 ratio in milk fatty acids was lower for SILc and LAT compared with LATc, indicating improved nutritional quality for SILc and LAT. However, retinol concentration in milk was lower in SILc compared with all other treatments. The study implies that feeding silage instead of fresh grass has no effect on DMI, ECM yield, or CH4 yield, and that concentrate supplementation can increase milk production, affects milk quality, and reduces the effect on climate, whereas feeding less mature grass increases DMI and milk yield, but has no effect on CH4 yield.

Correlation of attention deficit hyperactivity disorder with gut microbiota according to the dietary intake of Korean elementary school students.

We investigated the impact of dietary patterns on the gut microbiota and concentration of short-chain fatty acids in the feces of Korean elementary school students. The dietary intake and ADHD assessment of 40 Korean elementary school students were analyzed using a dish-based semi-quantitative food frequency questionnaire. Analysis of gut microbiota and short-chain fatty acids composition were performed using the real-time polymerase chain reaction, metagenomics, and gas chromatography methods. The dietary patterns of participants were divided into four groups: healthy, processed food, fish and shellfish, and meat. The participants were also divided into two groups according to their ADHD scores: 0–30, control group; over 30, ADHD group. The ADHD score of the processed food group was significantly higher than that of the healthy group. The processed food and ADHD groups showed significantly higher abundance of harmful bacteria, such as the Enterobacter, Escherichia coli, and Clostridium strains, and markedly lower abundance of beneficial bacteria, such as the Bifidobacterium and Ruminococcus strains, than the control group. The heat maps of metagenomics indicated that each group was separated into distinct clusters, and the processed food and ADHD groups showed significantly lower α-diversity of gut microbiota than the control group. In these groups, the concentration of acetate or butyrate in the feces was significantly lower than that in the control group. These results may indicate that imbalanced diets can disturb the colonic microbial balance and are likely to become a potential risk factor for the prevalence of ADHD.

Chronic dietary exposure to a glyphosate-based herbicide results in total or partial reversibility of plasma oxidative stress, cecal microbiota abundance and short-chain fatty acid composition in broiler hens.

Glyphosate-based herbicides (GBHs) are massively used in agriculture. However, few studies have investigated the effects of glyphosate-based herbicides on avian species although they are largely exposed via their food. Here, we investigated the potential reversibility of the effects of chronic dietary exposure to glyphosate-based herbicides in broiler hens. For 42 days, we exposed 32-week-old hens to glyphosate-based herbicides via their food (47 mg/kg/day glyphosate equivalent, glyphosate-based herbicides, n = 75) corresponding to half glyphosate’s no-observed-adverse-effect-level in birds. We compared their performance to that of 75 control animals (CT). Both groups (glyphosate-based herbicides and control animals) were then fed for 28 additional days without glyphosate-based herbicides exposure (Ex-glyphosate-based herbicides and Ex-control animals). Glyphosate-based herbicides temporarily increased the plasma glyphosate and AMPA (aminomethylphosphonic acid) concentrations. Glyphosate and aminomethylphosphonic acid mostly accumulated in the liver and to a lesser extent in the leg muscle and abdominal adipose tissue. Glyphosate-based herbicides also temporarily increased the gizzard weight and plasma oxidative stress monitored by TBARS (thiobarbituric acid reactive substances). Glyphosate-based herbicides temporarily decreased the cecal concentrations of propionate, isobutyrate and propionate but acetate and valerate were durably reduced. The cecal microbiome was also durably affected since glyphosate-based herbicides inhibited Barnesiella and favored Alloprevotella. Body weight, fattening, food intake and feeding behavior as well as plasma lipid and uric acid were unaffected by glyphosate-based herbicides. Taken together, our results show possible disturbances of the cecal microbiota associated with plasma oxidative stress and accumulation of glyphosate in metabolic tissues in response to dietary glyphosate-based herbicides exposure in broiler hens. Luckily, glyphosate-based herbicides at this concentration does not hamper growth and most of the effects on the phenotypes are reversible.

Orlistat and ezetimibe could differently alleviate the high-fat diet-induced obesity phenotype by modulating the gut microbiota.

This study aimed to evaluate the possible anti-obesity effects of orlistat and ezetimibe and determine the mechanism by which they alter the composition of gut microbiota and short-chain fatty acids (SCFAs) in mice with a high-fat diet (HFD)-induced obesity. Eighty male, specific pathogen-free C57BL/6J mice aged 3 weeks were divided into four groups (n = 20). The NCD group was fed with a normal diet, and the HFD, HFD+ORL, and HFD+EZE groups were fed with HFD for 20 weeks. From the 13th week onward, the HFD+ORL and HFD+EZE groups were administered with orlistat and ezetimibe, respectively. The glucose and lipid metabolism of the tested mice were evaluated by analyzing blood biochemical indicators during the intervention. Furthermore, the changes in the structure of the fecal microbiota and the fecal SCFA content were analyzed by 16S rRNA sequencing and gas chromatography-mass spectrometry, respectively. HFD induced the obesity phenotype in mice. Compared to the HFD group, the body weight, visceral fat-to-body weight ratio, serum total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C), and oral glucose tolerance test (OGTT) of the HFD+ORL group significantly decreased, whereas fecal butyric acid levels significantly increased. Ezetimibe intervention significantly reduced the OGTT, serum TC, and HDL-C levels only. The α-diversity of the gut microbiota significantly decreased after intervention with orlistat and ezetimibe. Orlistat altered the relative abundance of some bacteria in the fecal microbiota. The populations of Firmicutes, Alistipes, and Desulfovibrio decreased, whereas those of Verrucomicrobia and Akkermansia significantly increased. Ezetimibe caused changes only in some low-abundance bacteria, as manifested by a decrease in Proteobacteria and Desulfovibrio, and an increase in Bacteroides. The administration of orlistat and ezetimibe can characteristically influence the body weight and serum lipid metabolism, and glucolipid levels in diet-induced obese mice and is accompanied by significant changes in the gut microbiota and SCFAs. These results suggest that the two drugs might exert their own specific anti-obesity effects by modulating the gut microbiota in a different manner. The enhanced health-promoting effect of orlistat might result from its stronger ability to alter the gut microbiota and SCFAs, at least partly.

Alleviating effects of gut micro-ecologically regulatory treatments on mice with constipation.

Treatments targeted for gut microbial regulation are newly developed strategies in constipation management. In this study, the alleviating effects of gut micro-ecologically regulatory treatments on constipation in mice were investigated. Male BALB/c mice were treated with loperamide to induce constipation, and then the corresponding intervention was administered in each group, respectively. The results showed that administration of mixed probiotics (MP), a 5-fold dose of postbiotics (P5), both synbiotics (S and S2), as well as mixed probiotics and postbiotics (MPP) blend for 8 days shortened the time to the first black stool, raised fecal water content, promoted intestinal motility, and increased serum motilin level in loperamide-treated mice. Furthermore, these treatments altered gut microbial composition and metabolism of short-chain fatty acids (SCFA). Based on linear regression analysis, SCFA was positively correlated with serum motilin except for isobutyrate. It suggested gut microbial metabolites affected secretion of motilin to increase gastrointestinal movement and transportation function and thus improved pathological symptoms of mice with constipation. In conclusion, the alteration of gut micro-ecology is closely associated with gastrointestinal function, and it is an effective way to improve constipation via probiotic, prebiotic, and postbiotic treatment.

Dietary Protected Butyrate Supplementation of Broilers Modulates Intestinal Tight Junction Proteins and Stimulates Endogenous Production of Short Chain Fatty Acids in the Caecum.

Simple SummaryThe aim of this study was to evaluate the effect of protected butyrate supplementation in broiler nutrition. Two experiments were conducted: (i) a performance study (1–35 days of age) and (ii) a study on the gut integrity and microbiome (1–21 days of age). The growth performance of birds, abundance of intestinal tight junction proteins, the quantitative characteristics of caecal microbiome and intestinal short chain fatty acid (SCFA) concentrations were investigated in these studies. Based on the results, it could be concluded that the addition of protected butyrate to broiler diets improved growth performance and intestinal integrity in accordance with the modulation of the caecal microbiome and the stimulated endogenous production of SCFA in the caecum. These findings provide important information that increases the understanding of the complex effects of the biologically most active SCFA, butyrate, on the overall health status and metabolism of the chicken.Short chain fatty acid (SCFA) butyrate has various beneficial effects on the gut microbiota as well as on the overall health status and metabolism of the host organism. The modulatory role of butyrate on gut barrier integrity reflected by tight junction protein expression has been already described in mammalian species. However, there is limited information available regarding chickens. Therefore, the main aim of this study was to monitor the effects of protected butyrate on claudin barrier protein and monocarboxylate transporter 1 abundance in different gastrointestinal segments of chickens as well as the growth performance of broiler chickens. The effect of protected butyrate on the caecal microbiota was monitored by quantifying the concentrations of total eubacteria and key enzymes of butyrate production. Furthermore, intestinal SCFA concentrations were also measured. Based on the data obtained, protected butyrate increased the overall performance as well as the barrier integrity of various gut segments. Protected butyrate also positively affected the SCFA concentration and composition. These findings provide valuable insight into the complex effects of protected butyrate on broiler gut health, highlighting the beneficial effects in improving intestinal barrier integrity and performance parameters.

Acidogenic fermentation of food waste in a leachate bed reactor (LBR) at high volumetric organic Loading: Effect of granular activated carbon (GAC) and sequential enrichment of inoculum

This study investigated the impact of different granular activated carbon (GAC) loadings and inoculum enrichment on acidogenic fermentation of food waste in a leachate bed reactor (LBR) operated at a high volumetric organic loading of 49 g VS/Lreactor. LBR with a high GAC loading of 0.51 g GAC/g VSfood waste achieved hydrolysis yield of 620 g SCOD/kg VSadded, significantly (P ≤ 0.05) higher to that obtained for LBRs with low or no GAC loading. A high GAC loading also resulted in a higher acidification yield of 507 g CODSCFA/kg VSadded. Butyrate dominated the short-chain fatty acid (SCFA) composition by constituting 57–60 % of total SCFA at high GAC loadings, while the composition of acetate (38–40 %) and butyrate (36–38 %) were similar at lower GAC loadings. Inoculum enrichment further improved the hydrolysis and acidogenesis yields by 10–22 % resulting in the final hydrolysis yield of 683 g SCOD/kg VSadded and acidification yield of 617 g CODSCFA/kg VSadded.

Comparing a single dose of xylanase to a double dose or cocktail of non-starch polysaccharide-degrading enzymes in broiler chicken diets

This study compared supplementation with a single dose of xylanase to a double dose of xylanase or a non-starch polysaccharide (NSP) degrading enzyme cocktail (NSP-ase cocktail) on productive performance, nutrient utilisation and the gastrointestinal environment in broilers fed commercial diets. Cobb 500 broilers (n=1,080) were fed 12 dietary treatments; four Australian commercial diets (based on wheat-barley, wheat-maize, wheat-sorghum or wheat only) with three different enzyme treatments (single dose of xylanase (16,000 BXU/kg), double dose of xylanase (32,000 BXU/kg) or NSP-ase cocktail (xylanase, β-glucanase, cellulase, pectinase, mannanase, galactanase, arabinofuranosidase). There were 108 pens, nine replicates per dietary treatment, with 10 birds per pen. Performance (total pen body weight, feed intake and feed conversion ratio corrected for mortality) was determined at d 0-35. On d 35, one male and one female were weighed individually and used to determine breast meat, thigh and drumstick weight, dry matter (DM) contents from the gizzard, jejunum and ileum, ileal protein, energy, starch and dry matter digestibility, ileal viscosity and xylo-oligosaccharide (XOS) concentration, caecal microbiota and short chain fatty acid (SCFA) composition. The double dose of xylanase and NSP-ase cocktail had no effect on bird performance, meat yield, ileal viscosity, ileal starch, energy or DM digestibility or digesta DM content. The double xylanase dose and NSP-ase cocktail increased protein digestibility in birds fed the wheat-sorghum based diet (P=0.041) and increased caecal concentration of butyric acid in birds fed the wheat-maize based diet (P=0.040), and propionic, valeric and lactic acid and Bifidobacteria and Enterobacteria spp. in birds fed the wheat-based diet (P&lt;0.05). The double xylanase dose increased XOS production, particularly in birds fed the wheat-barley based diets (P&lt;0.05). The lack of performance effects observed when feeding the double xylanase dose or NSP-ase cocktail suggested that the current recommended xylanase dose (16,000 BXU/kg) is sufficient.

Effects of Oats, Tartary Buckwheat, and Foxtail Millet Supplementation on Lipid Metabolism, Oxido-Inflammatory Responses, Gut Microbiota, and Colonic SCFA Composition in High-Fat Diet Fed Rats.

Coarse cereals rich in polyphenols, dietary fiber, and other functional components exert multiple health benefits. We investigated the effects of cooked oats, tartary buckwheat, and foxtail millet on lipid profile, oxido-inflammatory responses, gut microbiota, and colonic short-chain fatty acids composition in high-fat diet (HFD) fed rats. Rats were fed with a basal diet, HFD, oats diet (22% oat in HFD), tartary buckwheat diet (22% tartary buckwheat in HFD), and foxtail millet diet (22% foxtail millet in HFD) for 12 weeks. Results demonstrated that oats and tartary buckwheat attenuated oxidative stress and inflammatory responses in serum, and significantly increased the relative abundance of Lactobacillus and Romboutsia in colonic digesta. Spearman’s correlation analysis revealed that the changed bacteria were strongly correlated with oxidative stress and inflammation-related parameters. The concentration of the butyrate level was elevated by 2.16-fold after oats supplementation. In addition, oats and tartary buckwheat significantly downregulated the expression of sterol regulatory element-binding protein 2 and peroxisome proliferator-activated receptors γ in liver tissue. In summary, our results suggested that oats and tartary buckwheat could modulate gut microbiota composition, improve lipid metabolism, and decrease oxidative stress and inflammatory responses in HFD fed rats. The present work could provide scientific evidence for developing coarse cereals-based functional food for preventing hyperlipidemia.

Astaxanthin attenuated the stress-induced intestinal motility disorder via altering the gut microbiota.

Gut microbiota and short-chain fatty acids (SCFAs) are recognized as key factors in the pathophysiology of irritable bowel syndrome. Astaxanthin is a carotenoid with strong antioxidant and anti-inflammatory activities. In this study, we examined the effects of astaxanthin on gut microbiota-, SCFAs-, and corticotropin-releasing factor (CRH)-induced intestinal hypermotility. Male Wistar rats (n=12 per group) were fed a diet with or without 0. 02% (w/w) astaxanthin for four weeks and CRH or saline was administered intravenously. The number of fecal pellets was counted 2 h after injection. Then the rats were sacrificed, and the cecal content were collected 3 h after injection. The number of feces was significantly increased by CRH injection in the control group (2.0 vs. 6.5; p=0.028), but not in the astaxanthin group (1.0 vs. 2.2; p=0.229) (n=6 per group). The cecal microbiota in the astaxanthin group was significantly altered compared with that in the control group. The concentrations of acetic acid (81.1 μmol/g vs. 103.9 μmol/g; p=0.015) and butyric acid (13.4 μmol/g vs. 39.2 μmol/g; p<0.001) in the astaxanthin group were significantly lower than that in the control group (n=12 per group). Astaxanthin attenuates CRH-induced intestinal hypermotility and alters the composition of gut microbiota and SCFAs.

Ameliorative effect of tuna elastin peptides on AIA mice by regulating the composition of intestinal microorganisms and SCFAs

• Tuna elastin peptides relieved adjuvant-induced arthritis in mice. • Tuna elastin peptides modulated gut microbiota and SCFAs metabolite profile. • Tuna elastin peptides induced a dose-dependent pattern in the AIA model. • The relieving arthritis of tuna elastin peptides were transmissible by FMT. Elastin peptides display a wide range of biological activities. This study aimed to investigate the ameliorative effect of tuna elastin peptides on adjuvant-induced arthritis (AIA). The daily administration of tuna elastin peptides markedly downregulated the levels of proinflammatory cytokines and upregulated the immunosuppressive cytokines in the serum, bone, and colon of the AIA model. 16S rRNA sequencing showed that tuna elastin peptide-treated mice exhibited reversed gut microbiota dysbiosis and increased abundance of Lactobacillus and Clostridium . Additionally, metabolomics indicated that short-chain fatty acid (SCFAs) production was increased in tuna elastin peptide-treated mice. The ameliorating arthritis effects of tuna elastin peptides were transmitted by transplanting the fecal microbiota from mice treated with high levels of elastin peptides. These results indicate the potential of tuna elastin peptides as beneficial functional food that ameliorates rheumatoid arthritis.

A Cocoa Diet Can Partially Attenuate the Alterations in Microbiota and Mucosal Immunity Induced by a Single Session of Intensive Exercise in Rats.

BackgroundFollowing intensive sports events, a higher rate of upper respiratory tract infections and the appearance of gastrointestinal symptomatology have been reported. We aimed to evaluate the effect of a cocoa-enriched diet on the cecal microbiota and mucosal immune system of rats submitted to high-intensity acute exercise, as well as to elucidate the involvement of cocoa fiber in such effects.MethodsWistar rats were fed either a standard diet, a diet containing 10% cocoa providing 5% fiber and a diet containing only 5% cocoa fiber. After 25 days, half of the rats of each diet performed an exhaustion running test. Sixteen hours later, samples were obtained to assess, among others, the cecal microbiota and short chain fatty acids (SCFAs) composition, mesenteric lymph nodes (MLNs) and Peyer’s patches (PPs) lymphocyte composition, and immunoglobulin (Ig) content in salivary glands.ResultsThe intake of cocoa, partially due to its fiber content, improved the SCFA production, prevented some changes in PPs and in MLNs lymphocyte composition and also decreased the production of proinflammatory cytokines. Cocoa diet, contrary to cocoa fiber, did not prevent the lower salivary IgM induced by exercise.ConclusionA cocoa dietary intake can partially attenuate the alterations in microbiota and mucosal immunity induced by a single session of intensive exercise.

Effects of Pathogenic Escherichia coli Infection on the Flora Composition, Function, and Content of Short-Chain Fatty Acids in Calf Feces.

Simple SummaryCalf diarrhea caused by bacteria usually begins to occur 2–3 d after birth. The main pathogenic bacteria are Escherichia coli, Campylobacter, Salmonella, and Clostridium perfringens. Pathogenic E. coli is the main bacterium that causes diarrhea in calves. It invades the body of calves and releases enteric toxins that cause diarrhea. The incidence of calf diarrhea caused by pathogenic E. coli O1 can reach 60–70%, even as high as 90–100% in large-scale calf farms, and the mortality rate can reach over 50%. Therefore, the prevention and treatment of calf diarrhea are extremely important. We established a calf diarrhea model by artificially intervening in the colonization of intestinal microbial flora and studied the influence of pathogenic E. coli on the composition, function, and short-chain fatty acid content of calf feces. Our research shows that pathogenic E. coli O1 can significantly reduce the abundance and diversity of calf intestinal microflora, increase harmful bacteria, reduce the colonization of beneficial bacteria, and reduce the short-chain fatty acid content in feces.Calf diarrhea caused by pathogenic Escherichia coli is a major cause of death in calves, with a mortality rate of over 50%. It is crucial to understand the pathogenesis and development of calf diarrhea for its prevention and treatment. We aimed to study the effect of pathogenic E. coli on the flora composition, function, and short-chain fatty acid (SCFA) content of calf feces using a calf diarrhea model. Sixty-four newborn Holstein calves (40–43 kg) were divided into a normal group (NG; n = 32) and a test group (TG; n = 32). At the beginning of the experiment, the TG were orally administered pathogenic E. coli O1 (2.5 × 1011 CFU/mL, 100 mL) to establish a calf diarrhea model, and the NG were orally administered the same amount of physiological saline solution. The calves of the two groups were subjected to the same feeding and management. Fresh feces samples were collected at different time points and subjected to 16S rRNA high-throughput sequencing and gas chromatography–mass spectrometry to determine the fecal microbial composition and SCFA content. Pathogenic E. coli O1 significantly altered microbiotas composition in the feces of calves, increasing the relative abundance of Proteobacteria and decreasing that of Firmicutes. It also led to a significant increase in the relative abundance of Escherichia-Shigella and a decrease in Lactobacillus, as well as significantly decreased SCFA content. Therefore, we postulate that pathogenic E. coli induces calf diarrhea by causing intestinal florae imbalance and reducing the content of SCFA.

A50 IMPACT OF SHORT CHAIN FATTY ACIDS ON PATHOGENICTY OF COMMENSAL BACTERIA IN PEDIATRIC INFLAMMATORY BOWEL DISEASES

Abstract Background Dietary fibers are fermented by microbiota into many different compounds including short-chain fatty acids (SCFA). The gut microenvironment is essential not only for human health but also in maintaining a rich biodiversity of bacterial species. It is well known that the microbiome is altered in Inflammatory Bowel Diseases (IBD), which likely changes the microenvironment and has other health effects on individuals with IBD. Pathobionts are organisms that can induce pathology under specific conditions, and several have already been associated with IBD, such as adherent invasive Escherichia coli (AIEC) and Helicobacter hepaticus. Although these bacteria have been associated with IBD, it is still largely unknown what conditions make these bacteria become pathogenic. Aims Determine the role of SCFA composition on the pathogenic potential of commensal bacteria isolated from non-IBD and IBD pediatric patients. Methods Anaerobic bacteria were isolated during colonoscopy from pediatric IBD and non-IBD patients and identified using 16S sequencing. PMA-activated human monocytes (THP-1) and colonic epithelial cells (Caco2) were grown in the presence of SCFA (acetate, butyrate, formate, propionate, and succinate). The ability of commensal bacteria ( Bacteroides fragilis, Clostridium innocuum, Ruminococcus, Parabacteroides merdae, Bifidobacterium infantis and E. coli HB101) and the pathobiont AIEC to invade cells was assessed using a gentamicin protection assay. Immune activation was quantified with IL-1b and IL-6 by ELISA, as well as reactive oxygen species (ROS) using DCFDHA. Results The SCFA propionate and formate significantly decreased invasion for HB101 and LF82 into macrophages but not epithelial cells. Bacteria isolated from an IBD environment had a higher invasion potential independent of the addition of SCFA, compared to bacteria from a non-IBD environment. However, butyrate was found to significantly increase invasion of non-IBD B. fragilis into Caco2 cells but not macrophages. Bacteria isolated from IBD patients tended to have a higher inflammatory response with both elevated IL-1b as well as ROS production. Conclusions Microenvironment changes, such as in SCFA concentrations, during a disease state can affect the host response to microbes including commensals. Commensal bacteria from IBD patients tend to be more proinflammatory, suggesting that they play a role in driving inflammation. SCFA are beneficial factors for gut health; however, what is still unknown is the interaction between microbes and the host after microenvironmental shifts that likely causes bacterial stress. Understanding the host-microbe interactions and the role diet can have on this dynamic relationship has the potential for discovering new therapeutic options for those suffering from IBD. Funding Agencies CIHRWeston Family Foundation

UniqPy: A tool for estimation of short-chain fatty acids composition by gas-chromatography/mass-spectrometry with headspace extraction

Short-chain fatty acids are metabolites widely presented in many natural sources, including human feces and blood. Estimation of their composition is a common procedure, usually performed using nuclear magnetic resonance or gas chromatography with a flame ionization detector. However, the commonly used methods often depend on specific sample preparation, such as filtration and homogenization. The gas-chromatography/mass-spectrometry (GC/MS) method with headspace extraction allows sample preparation to be kept to a minimum regardless of the physical state of the sample, which can be potentially useful in metabolomics research of complex natural samples such as blood or feces. In this work, we have demonstrated the applicability of Headspace GC-MS for estimating short chain fatty acid (SCFA) composition. The main problem here is the complex, non-linear dependence between the composition of the compounds in the source phase and the relative pressures in the vapor phase, which are directly measured by this method. We have implemented a thermodynamic model that performs the reverse transformation of relative abundances in the vapor phase to relative concentrations in the liquid phase, and have tested it on some synthetic SCFA mixtures. The developed method is available as a pip package called UniqPy and can be used to describe liquid-vapor equilibrium for any multicomponent system if a sufficient amount of training data is provided. The gas chromatography method with headspace extraction in conjunction with the UniqPy data transformation showed satisfactory quantification accuracy for propionic acid, butyric acid, isobutyric acid, and valeric acid (R-squared > 0.96). The applicability of the method was additionally demonstrated on a series of fecal samples.

CLA improves the lipo-nutritional quality of pork and regulates the gut microbiota in Heigai pigs.

Conjugated linoleic acid (CLA) is a potential nutritional strategy to regulate meat quality in pigs and produce high-quality pork. However, the effects of CLA on nutritional quality, lipid dynamics, microbiota, and their metabolites in the gut of pigs remain unclear. Our study explored the effects of CLA on lipo-nutritional quality based on a Heigai pig model and investigated the regulatory mechanism using an integrated analysis of multiple omics. A total of 58 Heigai finishing pigs (body weight: 85.58 ± 10.39 kg) were randomly divided into 2 treatments and fed diets containing 1% soyabean oil and 1% CLA for 40 days. 1% CLA significantly decreased the backfat thickness (P < 0.05) and increased the intramuscular fat (IMF) content (P < 0.05). The expression of lipid metabolism-related genes was significantly changed (P < 0.05) and lipidome analysis showed the alternations of lipid dynamics in the longissimus dorsi muscle (LDM). In addition, based on the microbiome and metabolomic analyses, the relative abundances of Parabacteroides, Bacteroides, and Lachnospiraceae_UCG-010 increased and CLA changed the metabolome profiles and the short-chain fatty acid (SCFA) composition in the gut, which were significantly increased (P < 0.05). Additionally, Pearson's correlation analysis indicated that differential microbial genera and SCFAs induced by CLA had tight correlations with the backfat thickness, IMF content and lipids in the LDM. CLA enhances the lipid accumulation and metabolism in muscle and these changes are associated with the production and functions of the differential bacteria and SCFAs in the gut of pigs.