Cecal Short-chain Fatty Acids Research Articles

Effects of Edible Grass (Rumex patientia L. × Rumex tianschanicus A. LOS) Leaf Powder on Growth Performance, Antioxidant Properties, Cecal Short-Chain Fatty Acids, and Microbial Community Levels in Broilers

The hybrid plant edible grass (Rumex patientia L. × Rumex tianschanicus A. LOS), a member of the Rumex genus, presents a novel food source with a protein content of approximately 30–40%. By incorporating non-traditional feed sources, such as edible grass leaf powder (EGLP), into broiler production, costs could be reduced. The experimental subjects of this study were Arbor Acres (AA) broilers. A total of 300 newly hatched broilers were randomly divided into to five groups, with each group consisting of six cages, housing 10 birds per cage. The control group was fed a basal diet composed of maize and soybean meal. In the experimental groups, varying percentages (3%, 6%, 9%, and 12%) of EGLP were substituted for the corn–soybean meal in the basal diet. In the diet, for days 1–21, the corn content was reduced from 1.90% to 8.20%, and the soybean meal content was lowered from 1.50% to 6.00%. Similarly, in the diet, for days 22–42, the corn content was decreased from 1.17% to 7.00%, while the soybean meal content was reduced by 1.50% to 6.00%. The experiment lasted 42 days and was divided into two phases: the brooding phase (days 1 to 21) and the finishing phase (days 22 to 42). The results show that substituting 3% and 6% EGLP for corn and soybean meal significantly increased the average daily gain (ADG) during the brooding period compared to the control group (p < 0.01). Additionally, the group with a 3% substitution rate exhibited a significant increase in the average daily feed intake (ADFI) during the brooding phase (p < 0.01). Throughout the 42-day experiment, substituting 3% and 6% of EGLP for maize and soybean meal did not significantly affect the overall growth performance of broilers (p > 0.05). However, a 6% supplementation notably reduced the feed conversion ratio (FCR) (p < 0.01). Both the 3% (EG3) EGLP and 6% (EG6) EGLP replacement meals significantly enhanced the antioxidant capacity of broiler chickens, as indicated by the increased levels of the total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) (p < 0.01). Additionally, broilers supplemented with 3–12% showed a marked increase in cecal short-chain fatty acids (SCFAs) compared to the control group (p < 0.01). The 3% EGLP replacement diet also significantly boosted the prevalence of Lactobacillus in the cecum (p < 0.01). Furthermore, after EGLP supplementation, there was a higher abundance of SCFA-metabolizing bacteria, particularly Alistipes and Christensenellaceae_R-7, compared to pre-treatment (p < 0.05). The prevalence of Clostridium was significantly greater in the groups receiving 9% and 12% EGLP (p < 0.05), while Butyrivibrio levels were notably higher after supplementation with 12% EGLP (p < 0.05).

Effects of Lagenaria siceraria (Molina) Standl polysaccharides on growth performance, immune function, cecum microorganisms and short-chain fatty acids in broilers.

In this study, Lagenaria siceraria (Molina) Standl polysaccharides (LSP) was prepared using the water-alcohol precipitation method to evaluate its effects on growth performance, slaughter performance, cytokines, immune organ indices, cecal short-chain fatty acids (SCFAs), and microbial community structure in broiler chickens when added to the basal diet. Seventy-five broiler chickens were selected and randomly divided into five groups, with 15 chickens per group. All groups were fed a basal diet for 7 days. From 7 days of age, the control group continued to receive the basal diet, while the positive drug group was fed a diet supplemented with Astragalus polysaccharides (APS, 100 g/kg) in addition to the basal diet. The experimental groups were fed diets containing different concentrations of LSP (50, 100, and 200 g/kg) in addition to the basal diet, and the supplementation continued for 42 days. The findings indicated that the incorporation of LSP into the feed significantly enhanced average daily weight gain (ADWG), average daily feed intake (ADFI), feed to gain ratio (F/G), dressing percentage, percentage of breast muscle, percentage of leg muscle, and percentage of abdominal fat while concurrently reducing drip loss rate and cooking loss rate (p < 0.01) in comparison to the control group. Additionally, it significantly augmented the levels of interleukin-4 (IL-4) and interleukin-12 (IL-12) in cytokines, secreted immunoglobulin A (SIgA) and immunoglobulin G (IgG) in immunoglobulins, as well as immune organ indicators (p < 0.05). Furthermore, LSP also modulated the intestinal microbiome composition by increasing the abundance of Bacteroides species and significantly changing concentrations of specific short-chain fatty acids (SCFAs) such as propionic acid, isobutyric acid, acetic acid, and isovaleric acid (p < 0.01). These results suggest that dietary supplementation with LSP can effectively regulate intestinal microbiome composition while promoting short-chain fatty acid production. The alterations in microbial characteristics ultimately contribute to improved intestinal immunity and immune organ development as well as enhanced production performance and immune function in broilers.

Bacillus licheniformis and phytase combination in broiler chicken diets: Effects on growth performance, digestibility coefficients, gut microbiome activities, and bone quality

In the present study, B. licheniformis was added to broiler chicken diets alone or in combination with 6-phytase and the effects on growth performance, coefficients of ileal nutrient digestibility, morphometrical measurements of selected internal organs, gut microbial activity, and bone quality were determined. Four hundred 1-day-old female birds (Ross 308) were randomly allotted to 4 dietary treatments (10 replicate pens, 10 birds each). The trial was set up as follows: negative control (NC), basal diet without any feed additive; NC+Pro, basal diet with B. licheniformis (500 g/t of diet); NC+Phy, basal diet with 6-phytase (400 g/t of diet); and NC+Pro+Phy, basal diet in which both experimental factors were added at the same levels as above. The interaction between treatments was noted in each evaluated period regarding BWG and FI. In the entire experiment, an increased body weight gain (P < 0.001) was observed in the NC+Pro+Phy treatment compared to the other groups. The birds that received both feed additives combined were characterized by an increased feed intake (P < 0.001), contrary to NC+Pro and NC+Phy. Moreover, in terms of microenvironment modulations, interactions (P < 0.05) were observed in the jejunal and cecal digesta pH, short-chain fatty acid (SCFA) level, and microbial enzyme activities. However, the impacts of B. licheniformis and phytase on cecal SCFA concentrations and bacterial enzymes were mainly opposite. There were no interactions (P < 0.05) in terms of geometrical, mechanical, and structural selected tibia and femur indices. However, 6-phytase addition improved bone structure, stiffness, and elasticity. In conclusion, the interactions between 6-phytase and B. licheniformis positively affect growth performance via microbial activity changes; however, the supportive bone quality maintenance properties are only connected to 6-phytase supplementation.

Effects of xylo-oligosaccharide supplementation on the production performance, intestinal morphology, cecal short-chain fatty acid levels, and gut microbiota of laying hens

This study investigated the effects of xylo-oligosaccharide supplementation on the production performance, intestinal morphology, cecal short-chain fatty acid levels, and gut microbiota of laying hens. A total of 800 Lohmann pink layers, each 48 wk old, were randomly divided into 5 dietary treatment groups, namely XOS at 0 (CON), 100 (XOS1), 200 (XOS2), 300 (XOS3) and 400 (X0S4) mg/kg. The experimental period was 24 wk. The results revealed that the egg production rate and the number of eggs laid by each layer between 1 to 12 wk increased as the XOS concentration increased (Plinear < 0.05). The sand-shell egg percentage decreased significantly from 1 to 12 wk in the XOS1, XOS2, and XOS3 groups (PANOVA < 0.05). Compared with the CON group, the 4 XOS dosage groups presented significant increases in the villus height and the ratio of villus height to crypt depth in the jejunum (PANOVA < 0.05), whereas a linear decrease in jejunal crypt depth (Plinear < 0.05) was noted. In addition, XOS supplementation significantly increased the concentrations of butyric acid and isovaleric acid in the caeca (PANOVA < 0.05). High-throughput sequencing analysis of bacterial 16S rRNA revealed that dietary XOS supplementation influenced the cecal microbiota. The alpha diversity analysis indicated that the richness of cecal bacteria was greater in the laying hens fed XOS. The modulation of the cecal microbiota composition upon the addition of XOS was characterized by an increased abundance of Firmicutes and Bifidobacteriaceae, and decreased abundance of Bacteroidetes. At the genus level, dietary XOS supplementation resulted in decreases in the abundances of Bacteroides and Rikenellaceae_RC9_gut_group and an increase in the abundance of Lactobacillus. In conclusion, dietary XOS supplementation improved the production performance of laying hens by increasing the production of short-chain fatty acids and improving their intestinal morphology, which was achieved mainly through changes in the composition of the intestinal microbiota. The recommended level of XOS in the diet of laying hens is 200 mg/kg.

Microbiota but not immune modulation by a pro- and postbiotic was associated with the diet-additive interaction in broilers

This study investigated the diet-additive interactions of a Lactobacilli-based probiotic (Pro) and postbiotic (Post) on immune parameters and cecal microbiota composition, with subsequent effects on the metabolome in broilers. A completely randomized block design was employed with 2 diets [standard (SD), and challenge (CD)] and 3 additive conditions (Control, Pro, Post) involving 1,368 one-day-old male Ross 308 broilers equally distributed among 36 pens in a 42 d study. Diets were formulated to contain identical nutrient levels, with CD higher than SD in non-starch polysaccharide content by including rye and barley. Total non-specific serum Ig A, M and G concentrations were determined weekly from d14 to 35. Following vaccination, titres of specific antibodies binding Newcastle disease virus (NDV) and infectious bursal disease virus (IBDV) were measured. Microbiota composition was analyzed by 16S rRNA gene sequencing at d14 and 35, and α- and β-diversity indexes (Observed, Chao1, Bray, Jaccard) were calculated. Cecal short-chain fatty acids and the semi-polar metabolome were determined in the Control SD and all CD groups at d35. At d35, a diet-additive interaction was observed on cecal microbiota composition. Within SD, Pro and Post did not affect operational taxonomic units (OTU) abundance (adjusted-P > 0.05) and diversity indexes (P > 0.05). Within CD, Pro and Post affected the relative abundances of 37 and 44 OTUs, respectively (adjusted-P < 0.05), with Post but not Pro affecting β-diversity indexes (P = 0.041 and 0.064 for Bray and Jaccard, respectively). Within CD, Post increased cecal acetate (21%; P = 0.007) and butyrate (41%; P = 0.002) concentration and affected the concentration of 2 metabolites (adjusted-P < 0.05), while Pro affected 240 metabolites (adjusted-P < 0.05). No diet-additive interactions were observed on serum Ig (P > 0.05), except for IgM at d14 (P = 0.004). Diet composition, but not the additives, affected immune status parameters. The Pro and Post affected cecal microbiota composition only under dietary challenging conditions as previously reported for growth.

Effects of Early Life Exposures to the Aryl Hydrocarbon Receptor Ligand TCDF on Gut Microbiota and Host Metabolic Homeostasis in C57BL/6J Mice.

Exposure to persistent organic pollutants (POPs) and disruptions in the gastrointestinal microbiota have been positively correlated with a predisposition to factors such as obesity, metabolic syndrome, and type 2 diabetes; however, it is unclear how the microbiome contributes to this relationship. This study aimed to explore the association between early life exposure to a potent aryl hydrocarbon receptor (AHR) agonist and persistent disruptions in the microbiota, leading to impaired metabolic homeostasis later in life. This study used metagenomics, nuclear magnetic resonance (NMR)- and mass spectrometry (MS)-based metabolomics, and biochemical assays to analyze the gut microbiome composition and function, as well as the physiological and metabolic effects of early life exposure to 2,3,7,8-tetrachlorodibenzofuran (TCDF) in conventional, germ-free (GF), and Ahr-null mice. The impact of TCDF on Akkermansia muciniphila (A. muciniphila) in vitro was assessed using optical density (OD 600), flow cytometry, transcriptomics, and MS-based metabolomics. TCDF-exposed mice exhibited lower abundances of A. muciniphila, lower levels of cecal short-chain fatty acids (SCFAs) and indole-3-lactic acid (ILA), as well as lower levels of the gut hormones glucagon-like peptide 1 (GLP-1) and peptide YY (PYY), findings suggestive of disruption in the gut microbiome community structure and function. Importantly, microbial and metabolic phenotypes associated with early life POP exposure were transferable to GF recipients in the absence of POP carry-over. In addition, AHR-independent interactions between POPs and the microbiota were observed, and they were significantly associated with growth, physiology, gene expression, and metabolic activity outcomes of A. muciniphila, supporting suppressed activity along the ILA pathway. These data obtained in a mouse model point to the complex effects of POPs on the host and microbiota, providing strong evidence that early life, short-term, and self-limiting POP exposure can adversely impact the microbiome, with effects persisting into later life with associated health implications. https://doi.org/10.1289/EHP13356.

Metabolic Effects of Synbiotic Supplementation in High Fat Diet Fed Rats: Possible Role of GLP-1 and Serotonin

Abstract High fat diet (HFD) predisposes to many metabolic changes; it may disrupt gut barrier integrity and gut microbiota composition. Synbiotic supplementation may promote host’s metabolic health by selective activation of the healthy microorganisms. This study aimed to probe the interaction between synbiotic supplementation, gut microbiota and GUT hormones in HFD states. Twenty-seven adult male albino rats, 3 groups, group I: control, group II: HFD received HFD for 12 weeks and group III: synbiotic-supplemented HFD received synbiotic in the last 6 weeks. The anthropometric measurments were measured. Liver transaminases, lipid profile, parameters of insulin resistance, serum serotonin, glucagon like polypeptide-1 (GLP-1), oxidant/antioxidant markers (MDA/GPx), zonulin levels and quantitative cecal short chain fatty acids (SCFA) were 3 assessed. Samples of liver and colon were employed for histopathological studies. Compared to HFD group, synbiotic led to a significant reduction in anthropometric measurements, liver enzymes, atherogenic index, HOMA-IR and MDA denoting improved dyslipidemia, insulin resistance and oxidative state. Moreover, synbiotic supplementation decreased serum zonulin and increased both serum serotonin, GLP-1 and cecal SCFAs. Synbiotic supplementation ameliorated the metabolic derangements and the disturbed integrity of the intestinal barrier induced by HFD through reducing serum GLP-1, serotonin and cecal SCFAs.

Dietary jack bean (Canavalia ensiformis L.) supplementation enhanced intestinal health by modulating intestinal integrity and immune responses of broiler chickens

This study investigated the influence of supplementing with jack beans on jejunal morphology, cecal short-chain fatty acids production, gene expression both of pro- and anti-inflammatory cytokines and tight junctions. Four treatment groups including 288 Indian River chicks that were one day old were randomized at random. While the treatment groups received jack bean supplementation at levels of 5 %, 10 %, and 15 %, the control group (0 %) was given a basal diet. For 11–35 days, each treatment consisted of 8 pens with 9 birds each. Supplementing with jack beans significantly enhanced butyrate production (P < 0.001), while at 10 % supplementation did not differ from control. Villus height (VH) and the ratio (VH:CD) were significantly (P < 0.001) increased by dietary treatments, while villus width (VW) and crypt depth (CD) were significantly (P < 0.05) decreased. TLR-3, TNF-a, and IL-6 were all significantly (P < 0.001) increased by dietary supplementation. However, at 15 %, TLR-3 and IL-6 were same with control. IL-18 was significantly (P < 0.05) decreased at 15 %. IL-10 decreased significantly (P < 0.001), but at 10 % same with control. At 5 and 10 %, IL-13 increased significantly (P < 0.001), whereas dietary treatments decreased at 15 % compared to control. Although ZO1 decreased significantly (P < 0.001) and OLCN increased significantly (P < 0.001), both ZO1 and OCLN were not significantly different from the control at 15 %. Dietary treatments significantly (P < 0.001) increased CLDN1 but did not differ from the control at 10 %. JAM2 decreased significantly (P < 0.001) with dietary treatments. In conclusion, jack bean supplementation may increase broiler chicken performance and intestinal health due to butyrate production. It may affect intestinal morphology and integrity by upregulating a tight junction protein gene. Jack beans also impacted jejunum immune responses and inflammatory cytokine gene expression.

Inclusion of slowly digestible starch source is a promising strategy than reducing starch to protein ratio in low protein broiler diets

The present study investigated the effects of low protein diets with different starch sources and starch to protein ratio on growth, digestibility, intestinal health, caecal short chain fatty acids (SCFAs), serum cholesterol and triglycerides in broiler chickens. Eight hundred one-day-old male broiler chicks (Ross 308) were randomly allotted to one of 4 dietary treatments with 10 repeats and 20 birds in each repeat. The dietary treatment included 1) a standard protein corn-SBM based diet (SP), 2) a low protein corn-SBM based diet (LPI) without reduced starch: protein ratio, 3) a low protein corn-SBM based diet (LPII) with reduced starch: protein ratio, and 4) a low protein corn-SBM-peas based diet (LPP) and reduced starch: protein ratio. Soy hulls were added in the LPII and LPP diets to reduce starch: protein ratio. During the experiment period from 11-24 d, FI was not affected by the dietary treatments (P > 0.05). The BWG was significantly reduced in the LPI diet compared to the SP diet (P < 0.05). Likewise, FCR deteriorated in LPI and LPII but was better in the SP diet followed by the LPP diet (P < 0.05). The apparent total tract digestibility (ATTD) of dry matter (DM) varied significantly among the dietary treatments (P < 0.01). While ATTD of starch was similar for all the diets except the LPP diet wherein the ATTD of starch was significantly lower (P < 0.001). Ether extract digestibility was also significantly different between the SP and LPII dietary treatments (P < 0.01). The AME and AMEn values were also significantly lower in the LPP diet compared with other dietary treatments (P < 0.001). Nitrogen retention (%) was increased in all the LP diets compared with the SP diet (P < 0.001), but it was significantly better in both LPII and LPP diets compared to the LPI diet. The data showed that cecal short chain fatty acids (SCFAs) production was increased in the LPII and LPP compared to the SP and LPI diets (P < 0.001). Further, the production of acetic, butyric, and propionic acids was substantially higher in the LPP diet (P < 0.001). There was no significant difference in gene expression of Claudin-1 and ZO-1 (P > 0.05). However, MUC-2 and GLUT-1 gene expression were significantly downregulated in the LPI diet (P < 0.05). The concentration of cholesterol and triglycerides was significantly increased in the LPI diet (P < 0.001). In conclusion, the addition of peas as a slowly digestible starch source combined with soy hulls in low protein diet helped to partly recover the growth performance and improved cecal SCFAs production compared to other low protein diets with and without reduced starch: protein ratio in broiler chickens.

Dietary Clostridium butyricum and 25-Hydroxyvitamin D3 modulate bone metabolism of broilers through the gut—brain axis

Leg disorders have become increasingly common in broilers, leading to lower meat quality and major economic losses. This study evaluated the effects of dietary supplementation with Clostridium butyricum (C. butyricum) and 25-hydroxyvitamin D3 (25-OH-D3) on bone development by comparing growth performance, tibial parameters, Ca and P contents of tibial ash, bone development-related indicators' level, and cecal short-chain fatty acids in Cobb broilers. All birds were divided into four treatment groups, which birds fed either a basal diet (Con), basal diet + 75 mg chlortetracycline/kg (Anti), basal diet + C. butyricum at 109 CFU/kg (Cb), basal diet + C. butyricum at 109 CFU/kg and 25-OH-D3 at 25 μg/kg (CbD), or basal diet + 25-OH-D3 at 25 μg/kg (CD). Our results suggest that the dietary supplementation in Cb, CbD, and CD significantly increased the body weight (BW) and average daily gain (ADG), and reduced the feed-to-weight ratio (F/G) at different stages of growth (P < 0.05). Dietary supplementation in Cb, CbD, and CD prolonged (P < 0.05) the behavioral responses latency-to-lie (LTL) time, reduced (P < 0.05) the levels of osteocalcin (BGP) and peptide tyrosine (PYY), and increased (P < 0.05) serotonin (5-HT) and dopamine (DA). Treatment with Cb increased (P < 0.05) the levels of acetic acid, isobutyric acid, butyric acid, and isovaleric acid compared with those in Con group. The cecal metagenome showed that Alistipes spp. were significantly more abundant in Cb, CbD, and CD groups (P < 0.05). A total of 12 metabolic pathways were significantly affected by supplementation, including the signaling pathways of glucagon, insulin, and PI3K-AKT; primary and secondary bile acid biosynthesis; and P-type Ca 2+ transporters (P < 0.05). Hence, the CbD supplementation modulates bone metabolism by regulating the mediators of gut-brain axis, which may inform strategies to prevent leg diseases and improve meat quality in broilers.

Characterization of a novel sulfated-rhamnoglucuronan isolated from Korean seaweed Ulva pertusa and its efficacy for treatment of inflammatory bowel disease in mice

This study aimed to isolate Ulva pertusa polysaccharide (UPP), which elicits anti-inflammatory bowel disease (IBD) effects, from the Korea seaweed U. pertusa and identify its structure. Firstly, UPP was isolated from U. pertusa using hydrothermal extraction and ethanol precipitation. UPP is a novel polysaccharide that exhibits unique structural features such as 3-sulfated rhamnose, glucuronic acid, iduronic acid, and 3-sulfated xylose, which are repeated in 1,4-glycosidic bonds. Prophylactic oral administration of UPP in mice with dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) suppressed the levels of inflammatory cytokines and MAPK- and NF-κB-related factors in the serum and colon tissue. Tight junction (TJ)-related factors such as occludin, claudin-1, and mucin were effectively augmented by UPP in the colon tissue. In addition, UPP administration prevented the DSS treatment-led cecal short chain fatty acid imbalance, and this effect was most evident for propionic acid. In conclusion, UPP isolated from the Korean U. pertusa demonstrates potent anti-IBD activity. Characterization of this ulvan revealed its unique structure. Moreover, its efficacy may be associated with its anti-inflammatory effects and regulation of gut microbiota and TJ proteins. Thus, this study provides new insights into the biological effects of UPP in IBD.

Response of laying hens fed diet supplemented with a mixture of olive, laurel, and rosemary leaf powders: Metabolic profile, oxidative status, intestinal histomorphology, and egg quality

This study aimed to evaluate the effects of a mixture of olive, laurel, and rosemary leaf powders, on the oxidative state, biochemical, immune, intestinal morphophysiological parameters, and egg quality of laying hens. One hundred Lohmann Brown hens (28 weeks old) were equally assigned to two groups (n. 50) corresponding to a basal control diet (CON) or the diet supplemented with 6 g/kg feed of leaf powder mixture (LPM) containing olive, laurel, and rosemary leaves (1:1:1), for 60 days. Oxidative status, biochemical indices, immune response, cecal short chain fatty acids (SCFAs), intestinal morphological characteristics, and some egg traits were evaluated at the end of the experiment. The results indicated that LPM improved (P < 0.05) the oxidative status (TOS, ROMs), the immune system (IL-6, IL-1β, and TNF-α), the total protein and HDL cholesterol content, whereas it decreased (P < 0.05) total cholesterol and LDL cholesterol. Aspartate aminotransferase (AST), alkaline phosphatase (ALP), and alanine aminotransferase were significantly (P < 0.05) lower in the LPM than in the CON group. A significant increase (P < 0.05) in SCFA content in the caecum, as well as in villi height and crypt depth in both duodenum and ileum of LPM-treated hens, was observed. Egg quality parameters were not influenced (P > 0.05) by LPM. These findings indicate that LPM can be considered a candidate as an antioxidant ingredient for functional food in laying hens.

Impact of Plant-Based Dietary Fibers on Metabolic Homeostasis in High-Fat Diet Mice via Alterations in the Gut Microbiota and Metabolites

BackgroundThe gut microbiota contributes to metabolic disease, and diet shapes the gut microbiota, emphasizing the need to better understand how diet impacts metabolic disease via gut microbiota alterations. Fiber intake is linked with improvements in metabolic homeostasis in rodents and humans, which is associated with changes in the gut microbiota. However, dietary fiber is extremely heterogeneous, and it is imperative to comprehensively analyze the impact of various plant-based fibers on metabolic homeostasis in an identical setting and compare the impact of alterations in the gut microbiota and bacterially derived metabolites from different fiber sources. ObjectivesThe objective of this study was to analyze the impact of different plant-based fibers (pectin, β-glucan, wheat dextrin, resistant starch, and cellulose as a control) on metabolic homeostasis through alterations in the gut microbiota and its metabolites in high-fat diet (HFD)-fed mice. MethodsHFD-fed mice were supplemented with 5 different fiber types (pectin, β-glucan, wheat dextrin, resistant starch, or cellulose as a control) at 10% (wt/wt) for 18 wk (n = 12/group), measuring body weight, adiposity, indirect calorimetry, glucose tolerance, and the gut microbiota and metabolites. ResultsOnly β-glucan supplementation during HFD-feeding decreased adiposity and body weight gain and improved glucose tolerance compared with HFD-cellulose, whereas all other fibers had no effect. This was associated with increased energy expenditure and locomotor activity in mice compared with HFD-cellulose. All fibers supplemented into an HFD uniquely shifted the intestinal microbiota and cecal short-chain fatty acids; however, only β-glucan supplementation increased cecal butyrate concentrations. Lastly, all fibers altered the small-intestinal microbiota and portal bile acid composition. ConclusionsThese findings demonstrate that β-glucan consumption is a promising dietary strategy for metabolic disease, possibly via increased energy expenditure through alterations in the gut microbiota and bacterial metabolites in mice.

Heat-killed Akkermansia muciniphila ameliorates allergic airway inflammation in mice.

Allergic asthma (AA) is a common inflammatory airway disease characterized by increased airway hyper-responsiveness (AHR), inflammation, and remodeling. Akkermansia muciniphila is a strictly anaerobic bacterium residing in the gut and is a promising next-generation probiotic to improve metabolic inflammatory syndrome. A recent study suggested the beneficial effect of live A. muciniphila on allergic airway inflammation (AAI) in mice. However, whether the heat-killed form can improve AAI requires further investigation. Mice sensitized and challenged with house dust mites (HDM) develop AA hallmarks including inflammatory cell infiltration, goblet cell hyperplasia, and subepithelial collagen deposition in the lungs. These phenomena were reversed by oral administration of the heat-killed A. muciniphila strain EB-AMDK19 (AMDK19-HK) isolated from the feces of healthy Koreans. Furthermore, AMDK19-HK diminished the HDM-induced AHR to inhaled methacholine, lung mast cell accumulation, and serum HDM-specific IgE levels. It also led to the overall suppression of IL-4, IL-13, and eotaxin production in bronchoalveolar lavage fluids, and Il4, Il5, Il13, and Ccl17 gene expression in lung tissues. Moreover, AMDK19-HK suppressed Th2-associated cytokine production in the splenocytes of HDM-sensitized mice in vitro. Additionally, a combination of 16S rRNA gene sequencing and short-chain fatty acid (SCFA) analysis in cecal samples revealed that AMDK19-HK modulated the relative abundance of circulating SCFA-associated gut genera, including a positive correlation with Lachnospiraceae_ NK4A136_group and a negative correlation with Lachnoclostridium and significantly increased cecal SCFA concentrations. Finally, AMDK19-HK improved intestinal mucosal barrier function. These results suggest that the oral administration of AMDK19-HK ameliorates HDM-induced AAI in mice by suppressing Th2-mediated immune responses and could have a protective effect against AA development.

Effect of in ovo feeding of xylobiose and xylotriose on plasma immunoglobulin, cecal metabolites production, microbial ecology, and metabolic pathways in broiler chickens

BackgroundDietary supplementation of xylooligosaccharides (XOS) has been found to influence gut health by manipulating cecal microbiota and producing microbe-origin metabolites. But no study investigated and compared the effect of in ovo feeding of xylobiose (XOS2) and xylotriose (XOS3) in chickens. This study investigated the effect of in ovo feeding of these XOS compounds on post-hatch gut health parameters in chickens. A total of 144 fertilized chicken eggs were divided into three groups: a) non-injected control (CON), b) XOS2, and c) XOS3. On the 17th embryonic day, the eggs of the XOS2 and XOS3 groups were injected with 3 mg of XOS2 and XOS3 diluted in 0.5 mL of 0.85% normal saline through the amniotic sac. After hatching, the chicks were raised for 21 d. Blood was collected on d 14 to measure plasma immunoglobulin. Cecal digesta were collected for measuring short-chain fatty acids (SCFA) on d 14 and 21, and for microbial ecology and microbial metabolic pathway analyses on d 7 and 21.ResultsThe results were considered significantly different at P < 0.05. ELISA quantified plasma IgA and IgG on d 14 chickens, revealing no differences among the treatments. Gas chromatography results showed no significant differences in the concentrations of cecal SCFAs on d 14 but significant differences on d 21. However, the SCFA concentrations were lower in the XOS3 than in the CON group on d 21. The cecal metagenomics data showed that the abundance of the family Clostridiaceae significantly decreased on d 7, and the abundance of the family Oscillospiraceae increased on d 21 in the XOS2 compared to the CON. There was a reduction in the relative abundance of genus Clostridium sensu stricto 1 in the XOS2 compared to the CON on d 7 and the genus Ruminococcus torques in both XOS2 and XOS3 groups compared to the CON on d 21. The XOS2 and XOS3 groups reduced the genes for chondroitin sulfate degradation I and L-histidine degradation I pathways, which contribute to improved gut health, respectively, in the microbiome on d 7. In contrast, on d 21, the XOS2 and XOS3 groups enriched the thiamin salvage II, L-isoleucine biosynthesis IV, and O-antigen building blocks biosynthesis (E. coli) pathways, which are indicative of improved gut health. Unlike the XOS3 and CON, the microbiome enriched the pathways associated with energy enhancement, including flavin biosynthesis I, sucrose degradation III, and Calvin-Benson-Bassham cycle pathways, in the XOS2 group on d 21.ConclusionIn ovo XOS2 and XOS3 feeding promoted beneficial bacterial growth and reduced harmful bacteria at the family and genus levels. The metagenomic-based microbial metabolic pathway profiling predicted a favorable change in the availability of cecal metabolites in the XOS2 and XOS3 groups. The modulation of microbiota and metabolic pathways suggests that in ovo XOS2 and XOS3 feeding improved gut health during the post-hatch period of broilers.

Gastrointestinal response to the early administration of antimicrobial agents in growing turkeys infected with Escherichia coli

This study investigated the effects of the early administration of enrofloxacin (E) or doxycycline (D) for the first 5 consecutive days of life, or the continuous administration of the coccidiostat monensin (M) throughout the rearing period on gastrointestinal function in turkeys infected with avian pathogenic Escherichia coli (APEC) in an early or later stage of rearing. Experiment 1 lasted 21 d, and turkeys in groups E, D, and M were infected with APEC on d 15. Experiment 2 lasted 56 d, and it had a factorial arrangement of treatments where birds in groups E, D, and M were infected with APEC on d 15 or d 50. In both experiments, control groups (C) consisted of infected and uninfected birds without antibiotic or coccidiostat administration. On d 21 (Experiment 1) and d 56 (Experiment 2), 8 birds from each subgroup were killed, and the ileal and cecal digesta were sampled to analyze the activity of bacterial enzymes and the concentrations of short-chain fatty acids (SCFA). The experimental treatments did not affect the final body weight or body weight gain of birds. Both experiments demonstrated that APEC contributed to an increase in ammonia levels of the cecal digesta (means from 2 experiments: 0.311 vs. 0.225 mg/g in uninfected birds) and ileal pH (6.79 vs. 6.00) and viscosity (2.43 vs. 1.83 mPa⋅s). Moreover, the E. coli challenge enhanced the extracellular activity of several cecal bacterial enzymes, especially in older turkeys infected with APEC in a later stage of life. The continuous administration of monensin throughout the rearing period resulted in a weaker gastrointestinal response in older birds, compared with the other 2 antibiotics administered for the first 5 d of life. The results of the study are inconclusive as both desirable and undesirable effects of preventive early short-term antibiotic therapy were observed in turkeys, including normalization of ileal viscosity and cecal ammonia concentration (positive effect), and disruption in cecal SCFA production (negative effect).

Abstract 2794: Consumption of dietary fiber correlates with a significant increase in a-diversity in the intestinal Microbiome of allo-HCT recipients and with lower GVHD-lethality in pre-clinical models

Abstract Background: Graft-versus-host disease (GVHD) is associated with a decrease in intestinal microbiota and beneficial short-chain fatty acids (SCFAs). Dietary fiber can modulate the intestinal microbiome and SCFA production which effect intestinal epithelium health and immunity. Therefore, we hypothesized that a high-fiber diet in recipients of an allogeneic hematopoietic cell transplantation (allo-HCT) could result in an increase of beneficial commensals, concentrations of SCFAs, and decreased risk of GVHD. Methods: We evaluated dietary data and fecal samples of allo-HCT patients receiving unmodified grafts (n=95) at MSKCC (day -10 to day +30 relative to allo-HCT) and correlated microbial α-diversity (16s rRNA, n=568 samples) with fiber intake and acute GVHD in No-GVHD (n=30), Non-LGI-GVHD (GVHD except lower gastrointestinal tract, n=41), and LGI-GVHD patients (lower gastrointestinal tract GVHD, n=24).We tested concentrations of cellulose fiber (0%, 6%, 12% and 40%) in a GVHD mouse model (C57BL/6J into BALB/c) to assess survival, microbial composition (shotgun sequencing n=180; n=30 per group), cecal SCFAs concentrations (GC-MS n=24, n=8 per group), and lastly, T cell subsets (Th1/Th17/Tregs) and colonic epithelial MHC-II expression (flow cytometry n=54, n=18 per group). Results: In allo-HCT patients, fiber intake correlated positively with microbial α-diversity (R=0.3, p=0.04), especially in the LGI-GVHD group (p=0.007). Additionally, patients with high fiber intake had higher microbial α-diversity compared to the low fiber cohort (p&amp;lt;0.0001).GVHD mice receiving a fiber-rich diet (12%), compared to 0% and 6%, had a higher microbial α-diversity (p=0.01); reduced GVHD lethality (p=0.02); and a decrease in the Enterococcus sp. (p=0.004) and Akkermansia sp. (p=0.01) abundance; taxa which are associated with worse overall survival and increased risk of lethal GVHD. In contrast, compared to 12%, the 40% fiber diet resulted in worse survival (p=0.002), lower α-diversity (p=0.05) and a higher abundance of Bifidobacterium sp. (p&amp;lt;0.0001).Cecal butyrate in GVHD mice was higher in response to the 12% vs. the 0% (p=0.01) and 6% (p=0.05) fiber diets. Mice treated with12% fiber displayed higher number of Tregs (CD4+CD25+FOXP3+), than those with 0% (p=0.001) and 6% (p=0.02); and a lower expression of MHC-II in epithelial cells (EpCAM+CD45-) vs. 0% fiber (p=0.02). Conclusions: We demonstrated that a higher fiber intake in allo-HCT patients is associated with an increase in α-diversity; especially in LGI-GVHD patients. In a pre-clinical GVHD-model, optimal fiber consumption (12% cellulose) leads to an increase in α-diversity, a decrease in pathogen relative abundance, increased T-regulatory cells, and a decrease in GVHD lethality. These results suggest that dietary fiber could be utilized to prevent GVHD. Citation Format: Jenny Paredes, Anqi Dai, Ruben J. Faustino Ramos, Peter Adintori, Teng Fei, Chi Nguyen, Harold Elias, Kristen Victor, Romina Ghale, Charlotte Pohl, Ana Gradissimo, Justin Cross, Oriana Miltiadous, Jonathan Peled, Marina Burgos da Silva, Marcel RM van den Brink. Consumption of dietary fiber correlates with a significant increase in a-diversity in the intestinal Microbiome of allo-HCT recipients and with lower GVHD-lethality in pre-clinical models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2794.

Solid‐state fermentation pro‐enzymes supplementation benefits growth performance, health, and intestinal microbiota of broiler chickens fed wheat‐based diet

AbstractWheat as a kind of diet material can be used for broiler production. However, due to non‐starch polysaccharides in wheat, wheat may lead to lower growth performance and worth health. To reverse the negative effect, solid‐state fermentation pro‐enzymes were added. In this experiment, growth performance, intestinal health‐related genes, short chain fatty acids (SCFAs) and intestinal microbiota were detected to find the effects of wheat meal and combined with enzymes on broiler chickens from 15 to 42 days of age. A total of 432 1‐day‐old Arbor Acres broiler chickens were fed corn‐based diet (CD) for 14 days as the preparation stage of the experiment. Then, they were randomly divided into three groups and fed three different kinds of diets which were corn‐based diet (CD group), wheat‐based diet (WD group), and SFP enzymes supplementation in WD (Enzymes+Wheat‐based diet group). The results showed that compared with broilers in CD group, broilers in WD group had lower weight gain and higher Feed conversion ratio (p &lt; 0.05) during the whole experimental period especially from day 15 to day 21, but there was no significant effect on feed intake (p &gt; 0.05). Moreover, SFP enzymes decreased the spleen index (p &lt; 0.05). Wheat also had trends to decrease the expression of ZO‐1 (p = 0.096) and increase the concentrations of acetate (p &lt; 0.05), butyrate (p &lt; 0.05) and total SCFAs (p &lt; 0.05), in which SFP enzymes caused the opposite results except for butyrate, and SFP enzymes even increased the expression of ZO‐1 (p &lt; 0.001) and OCCLUDIN (p = 0.075) and decreased the expression of TNF‐α (p &lt; 0.01). Meanwhile, wheat enhanced the abundances of Barnesiella and Bifidobacterium (p &lt; 0.05) and inhibited the abundances of Flavonifractor, Sellimonas, Lachnospiraceae_NK4A136_group, Subdoligranulum, and Ruminococcus_gauvreauii_group (p &lt; 0.05), and SFP enzymes could reverse the negative effects, and the changes in microbiota could explain the other different parameters. Collectively, wheat results in inflammation and worse growth performance, but SFP enzymes supplementation in WD benefits chickens' growth performance by improving intestinal barrier function, decreasing inflammation, modulating cecal microbiota and SCFAs production.

Dietary probiotic Lacticaseibacillus paracasei NSMJ56 modulates gut immunity and microbiota in laying hens

This study was performed to investigate supplementary effects of probiotic Lacticaseibacillus paracasei NSMJ56 strain on laying performance, egg quality, intestinal histology, antioxidant status, gut immunity and microbiota in laying hens. A total of ninety-six 21-wk-old Hy-Line Brown laying hens were randomly subjected to one of 2 dietary treatments: a control group fed a non-supplemented diet, or a probiotic group fed with a diet supplemented with 1 g of Lacticaseibacillus paracasei NSMJ56 (5 × 108 CFU/kg of diet). The trial lasted for 4 wk. Egg weight was increased (P < 0.05) in laying hens fed probiotic-fed diet compared with the control group. Dietary probiotics did not affect egg quality except for Haugh unit, which was improved (P < 0.05) in the probiotic-fed group. Neither jejunal histology nor cecal short-chain fatty acids were affected by dietary treatments. Dietary probiotics increased the activity of catalase compared with the control group. Flow cytometry analysis revealed that dietary probiotics elevated the CD4+ T cells, but not CD8+ T cells, in jejunal lamina propria. Based on the LEfSe analysis at the phylum and genus levels, Erysipelotrichales, Erysipelotrichia, Flintibater, Dielma, Hespellia, Coprobacter, Roseburia, Anaerotignum, and Coprococcus were enriched in the probiotic group compared with the control group. Taken together, our study showed that dietary probiotics could be used to improve some parameters associated with egg freshness and antioxidant capacity, and to partially alter T cell population and microbial community in laying hens.

Characterizing the Effect of Campylobacter jejuni Challenge on Growth Performance, Cecal Microbiota, and Cecal Short-Chain Fatty Acid Concentrations in Broilers.

This study aimed to understand the effect of C. jejuni challenge on the cecal microbiota and short-chain fatty acid (SCFA) concentration to form a better understanding of the host-pathogen interaction. Sixty broilers were randomly allocated into two treatments: control and challenge. Each treatment was replicated in six pens with five birds per pen. On day 21, birds in the challenge group were orally gavaged with 1 × 108C. jejuni/mL, while the control group was mock challenged with PBS. The C. jejuni challenge had no effect on body weight, feed intake, and feed conversion ratio compared to the control group. On day 28, the C. jejuni challenge decreased the observed features and Shannon index compared to the control group. On the species level, the C. jejuni challenge decreased (p = 0.02) the relative abundance of Sellimonas intestinalis on day 28 and increased (p = 0.04) the relative abundance of Faecalibacterium sp002160895 on day 35 compared to the control group. The C. jejuni challenge did not change the microbial function and the cecal concentrations of SCFA on days 28 and 35 compared to the control group. In conclusion, C. jejuni might alter the gut microbiota's composition and diversity without significantly compromising broilers' growth.