Jejunal Tissue Research Articles

Low feed intake at weaning reduces intestinal glutathione levels and promotes cysteine oxidation to taurine in pigs.

Weaning stress in pigs is associated with low feed intake and poor nutrient utilization. Cysteine is a sulfur amino acid with key roles in pig production, but how cysteine metabolism and requirements are affected by weaning stress should be better defined. The objective of this study was to determine the collective impact of weaning and feed restriction on tissue cysteine metabolism. Pigs were weaned at 21-d age without access to feed (W; 6.90 ± 0.81kg; n = 9; reflecting acute nutritional stress) or were not weaned and remained with the sow (NW; 6.81 ± 0.65kg; n = 8). At euthanasia (23-d age), blood, bile, and liver, jejunum, and ileum tissues were collected. Plasma, bile, and tissue amino acid and amino thiol concentrations were analyzed by HPLC. Activity of glutamate cysteine ligase (GCL) and glutathione synthetase (GSS), enzymes needed for glutathione (GSH) production, and cysteine dioxygenase 1 (CDO1) were determined with activity assays followed by HPLC analysis of reaction products. Plasma (271 versus 192 ± 19 µmol/L; P < 0.001) and liver (417 versus 298 ± 33 nmol/g; P < 0.05) Cys concentrations were increased in W compared to NW pigs. Despite greater plasma Cys, jejunum and ileum Cys content were not affected by weaning (P > 0.10), whereas γ-glutamylcysteine (γ-GlyCys), the immediate precursor of GSH, declined in both jejunum (14.3 versus 9.7 ± 1.4 nmol/g; P < 0.01) and ileum (11.2 versus 6.4 ± 0.8 nmol/g; P < 0.001) in W pigs. Glutathione content was lower in the jejunum (1379 versus 1720 ± 70 nmol/g; P < 0.05) and ileum (1497 versus 1740 ± 74 nmol/g; P < 0.05) in W pigs. In the jejunum, GCL activity tended to be greater (0.56 versus 0.39 ± 0.07 nmol γ-GluCys • mg-1 • min-1; P < 0.10), whereas GSS activity tended to be lower (1.11 versus 1.38 ± 0.10 nmol GSH • mg-1 • min-1; P < 0.10) in W compared to NW pigs. In the ileum, the activities of GCL and GSS were not affected by weaning (P > 0.10). Although liver CDO1 activity was not different between groups (P < 0.10), liver taurine was greater in W compared to NW pigs (5115 versus 2336 ± 912 nmol/g; P = 0.001). Bile concentrations of Cys (1203 versus 279 ± 103 µmol/L; P < 0.001) and cysteinylglycine (203 versus 117 ± 33 µmol/L; P < 0.10), the direct product of GSH degradation, were greater in W compared to NW pigs. Collectively, these results suggest that systemic Cys is not effectively utilized for gut GSH production in newly weaned pigs and is instead oxidized to taurine and eliminated in bile.

Neutrophil-activating protein in Bacillus spores inhibits casein allergy via TLR2 signaling

BackgroundMilk allergy commonly occurs in children, mainly caused by bovine-derived casein (CAS) protein. Neutrophil-activating protein (NAP) of Helicobacter pylori plays an immunomodulatory role with potential to suppress Th2-type immune responses. Bacillus subtilis (B. subtilis) spores are commonly used as oral vectors for drug delivery.ObjectiveTo investigate whether recombinantly expressed NAP on B. subtilis spores could be an effective treatment for CAS allergy in mouse.MethodsAfter CAS sensitization, mice were orally administered B. subtilis spores expressing recombinant NAP for 6 weeks. Allergic symptoms and parameters were evaluated after CAS challenge oral gavage, including allergic inflammation, splenic cytokines, and serum-specific antibodies. Protein levels of Toll-like receptor 2 (TLR2) and c-JUN in the jejunum tissue were measured by western blot. Bone marrow-derived macrophages (BMDMs) were stimulated with inactivated NAP spores to measure the influence on cytokine profiles in vitro.ResultsNAP recombinant spore treatment significantly reduced allergic symptoms and intestinal inflammation. Interleukin-12 and interferon-gamma levels increased, whereas serum CAS-specific IgG1 and IgE levels decreased. TLR2 and c-JUN expression levels were elevated in the jejunal tissue. Inactivated NAP spores polarized BMDMs to the M1 phenotype and enhanced cytokine expression, which were inhibited by a TLR2 neutralizing antibody.ConclusionNAP offers a new strategy in the treatment of CAS allergy by inhibiting the Th2 response, while eliciting macrophages to promote Th1 immune responses.

Efficacy and safety of Shengjiang Xiexin decoction on irinotecan-induced diarrhea in small cell lung cancer patients: a multicenter, randomized, double-blind, placebo-controlled trial

BackgroundIrinotecan is a standard chemotherapeutic agent in small cell lung cancer (SCLC), however, as a common adverse reaction, diarrhea limits the use of irinotecan. Shengjiang Xiexin decoction (SXD) has been used in various gastrointestinal diseases in China two thousand years ago. We designed this clinical trial to supply more evidences on the use of SXD as prophylaxis for irinotecan-induced diarrhea, especially for high-risk population predicted by gene testing of uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1).MethodsIn this clinical trial, 120 patients with SCLC were recruited from six hospitals in China. They received two cycles of chemotherapy, meanwhile they were randomized to receive SXD or placebo for 14 days of oral administration in each cycle of chemotherapy. The primary outcome is the incidence of diarrhea. And secondary outcomes include the the degree of diarrhea and neutropenia, the number of chemotherapy cycles with diarrhea, first occurrence time and duration of diarrhea. To evaluate the effect of SXD on the intestine, a rat model with delayed-onset diarrhea induced by irinotecan was established, and the expression of inflammatory factors including IL-1β, IL-6 and TNF-α, anti-inflammatory factors including IL-10, TGF- β in jejunal tissue was detected by ELISA.Results101 patients (53 in SXD group, 48 in placebo group) completed the trial. The incidence of diarrhea in SXD group and placebo group were 26.42% (14/53) and 52.08% (25/48), respectively (P < 0.05), and the degree of diarrhea also had significant differences (P < 0.05). In UGT1A1 high-risk population, the incidence of diarrhea in two groups were 9.09% and 66.67% (P < 0.05), but there was no significant differences in UGT1A1 low-risk population. The incidence of neutropenia with degree 1–3 between two groups was 20.75% vs 20.83%, 13.21% vs 18.57%, 9.43% vs 20.83% (P < 0.05). No severe adverse events were reported in any group. And animal studies had shown SXD reduced content of IL-1β, IL-6, TNF-α, increased content of IL-10, TGF-β in jejunum tissue.ConclusionsSXD had a prophylactic effect in the diarrhea induced by irinotecan, especially for UGT1A1 high-risk population, and this effect from SXD appeared to be maintained the completion of chemotherapy schedule. The mechanism of action of SXD was related to the regulation of inflammatory factors.Trial registration Chinese Clinical Trial Register: ChiCTR1800018490. Registered on 20 September 2018. https://www.chictr.org.cn/showproj.html?proj=25250. The preliminary protocol of this clinical study has been published in the journal “Trials” in the form of protocol before this paper (Deng et al. in Trials 21:370, 2020).

Undernutrition disrupts jejunal and ileal microbiota and epithelial tissue homeostasis in a pregnant sheep model

Nutrition consistently affects microbe-host interactions in the gastrointestinal tract. This study aimed to unravel how undernutrition reshapes the microbial composition and the homeostasis of epithelium in the jejunum and ileum. Sixteen late-gestation Hu-sheep were randomly assigned to the control group (n = 8, 100% ad libitum feeding levels) or the undernutrition group (n = 8, which received 30% ad libitum feeding levels). After 15-d treatment, all ewes were slaughtered, and jejunal and ileal digesta and epithelium samples were collected for 16S rRNA gene sequencing and transcriptome sequencing, respectively. Results indicated that undernutrition decreased the jejunal and ileal tissue weights (P = 0.005 and P = 0.022) and the levels of volatile fatty acids (P = 0.019 and P = 0.007) and microbial protein levels (P = 0.019 and P = 0.031) in jejunal and ileal digesta. The relative abundance of acetate producing microbiota, including Clostridia UCG-014 norank, Ruminococcus, [Ruminococcus] gauvreauii, and Lachnospiraceae_Blautia, were significantly reduced (P < 0.05) in the jejunum and ileum. Undernutrition up-regulated (P < 0.05) the expression of genes involved in amino acid synthesis and fatty acid oxidation, but down-regulated (P < 0.05) the expression of genes associated with amino acid degradation, fatty acid synthesis, and extracellular structures in jejunal and ileal epithelium. In the jejunal epithelium, genes associated with extracellular matrix-receptor interactions, cell growth, and immune response were down-regulated (P < 0.05) upon undernutrition. Taken together, undernutrition changed the microbial community in the jejunum and ileum, which altered the fermentation mode and the production of volatile fatty acids and microbial protein. These affected the energy and protein system in the epithelium and reprogrammed substance metabolism and extracellular structures, which probably further influenced cell growth and immune response. These insights provide a foundation for completely clarifying the crosstalk between small intestinal microbiota and the host.

Transcriptome analysis of jejunal mucosal tissue in breeder hens exposed to acute heat stress

Transcriptome analysis of jejunal mucosal tissue in breeder hens exposed to acute heat stress

The effects of compound probiotics on production performance, rumen fermentation and microbiota of Hu sheep.

Fungal probiotics have the potential as feed additives, but less has been explored in ruminant feed up to date. This study aimed to determine the effect of compound probiotics (CPs) with Aspergillus oryzae 1, Aspergillus oryzae 2 and Candida utilis on Hu sheep's growth performance, rumen fermentation and microbiota. A total of 120 male Hu sheep, aged 2 months and with the body weight of 16.95 ± 0.65 kg were divided into 4 groups. Each group consisted of 5 replicates, with 6 sheep per replicate. Group A was the control group fed with the basal diet. Group B, C and D was supplemented with the basal diet by adding 400, 800 and 1,200 grams per ton (g/t) CPs, respectively. The feeding trial lasted for 60 days after a 10-day adaptation period. The results showed that the average daily gain (ADG) of sheep in the CPs groups were significantly higher, the feed/gain were significantly lower than those in group A in the later stage and the overall period. The addition of CPs increased the economic benefit. The levels of CD4+ and the CD4+/CD8+ ratio in the CPs groups were higher than those in Group A. The levels of GSH, IgG, IL-2, IL-6, and IFN-γ in group C were significantly elevated compared with group A. Group B showed a significant increase in rumen NH3-N and cellulase activity. There was no difference in VFAs content between group A and group B, however, with the increasing addition of CPs, the butyric acid and isobutyric acid content tended to decrease. The rumen microbiota analysis indicated that the CPs addition increased the Firmicutes and Proteobacteria abundances, decreased the Bacteroidetes abundance. The correlation analysis showed that Prevotella was negatively correlated with ADG, and the addition of 400 CPs in group B reduced Prevotella's relative abundance, indicating CPs increased sheep growth by decreasing Prevotella abundance. The CPs addition reduced caspase-3, NF-κB and TNF-α expression in liver, jejunum and rumen tissues. In conclusion, the addition of CPs increased the sheep production performance, reduced inflammation, improved rumen and intestinal health. Considering the above points and economic benefits, the optimal addition of CPs as an additive for Hu sheep is 800 g/t.

Functional remodeling of gut microbiota and liver in laying hens as affected by fasting and refeeding after fasting.

Animals will experience energy deprivation processes such as moulting, clutching, migration and long-distance transportation under natural survival conditions and in production practices, and the body will trigger a series of adaptive metabolic changes during these processes. Fasting and refeeding after fasting can induce remodeling of nutrients and energy metabolism. This study aims to investigate the mechanisms by which the gut microbiota and liver of poultry respond to energy deprivation under specific conditions. Ninety 252-day-old laying hens were randomly divided into 3 groups: (1) fed ad libitum (control group); (2) fasted from day 13 to day 17 (fasting group); (3) fasted from day 1 to day 5, then refed on a specific feeding way (refeeding group). After that, the serum, liver, jejunum tissues, and cecum contents were sampled and sent for metabolome, transcriptome, morphology, and 16S rDNA sequencing analyses, respectively. Results showed that food deprivation not only observably decreased the body weight, liver index, and the villus height and villus/crypt ratio of jejunum, but also significantly changed the gut microbiota compositions, serum metabolic profiles, and the hepatic gene expression patterns of laying hens, whereas these changes were effectively reversed by the following refeeding operation. At the same time, metabolome combined transcriptome analysis revealed that both serum differential metabolites and hepatic differential expressed genes (DEGs) were consistently enriched in the lipid and amino metabolism pathways, and strong correlations were synchronously found between the differential metabolites and both of the differential gut microbial genera and DEGs, suggesting the crosstalks among gut, liver and their resulting serum metabolic products. The results suggested that the organism might coordinate to maintain metabolic homeostasis under energy deprivation through a combination of changes in gut microbial composition and hepatic gene expression.

Impacts of replacing soybean meal with processed soybean meal on intestinal health and growth of nursery pigs challenged with F18+ Escherichia coli.

This study aimed to investigate the impact of different level of soybean meal (SBM) replaced by soy protein concentrate on intestinal health and growth performance of nursery pigs under F18+ Escherichia. coli (E. coli). Forty-eight newly weaned pigs (6.6 ± 0.3 kg) were randomly allotted to 4 treatments arranged by 2 × 2 factors using randomized complete block design with initial body weight and sex as blocks. Two factors were F18+ E. coli challenge (0 or 2.1 × 1010 CFU) and the level of SBM (24% or 12% in phase 1 and 26% or 14% in phase 2). Pigs were fed for 25 d in 2 phases (phase 1 for 11 d and phase 2 for 14 d). At the end of study, all pigs were euthanized to collect jejunal mucosa and tissues to measure parameters related to intestinal health. Data were analyzed using a MIXED procedure in SAS. The F18+ E. coli challenge decreased (p<0.05) overall average daily gain (ADG) and average daily feed intake (ADFI) and decreased (p<0.05) gain to feed ratio on d 7 to 11. The High SBM tended to have a greater overall ADG (p=0.054) and ADFI (p=0.078) compared with low SBM under F18+ E. coli challenge, but not in unchallenged conditions. The F18+ E. coli challenge increased (p<0.05) fecal score on d 7 to 18. The tumor necrosis factor-alpha and interleukin-1β in jejunal mucosa were decreased (p<0.05) in High SBM treatments. The High SBM tended to increase (p=0.085) occludin expression in jejunum. High SBM increased crypt depth in jejunum under F18+ E. coli challenge, but not in unchallenged conditions (p<0.05). High SBM in nursery diets could alleviate the detrimental effects of F18+ E. coli challenge on growth performance of pigs under compared to low SBM inclusion, which might be attributed to decreased intestinal inflammation and improved intestinal integrity.

Dietary glycerides of valerate ameliorate diarrhea and impact intestinal physiology and serum biomarkers in weaned piglets infected with enterotoxigenic Escherichia coli F18.

In the commercial swine farm setting, the post-weaning period is a critical window during which piglets are highly susceptible to infection and enterotoxigenic E. coli (ETEC)-associated diarrhea. Short chain fatty acids and their glycerides are compounds which may influence intestinal health; however, valerate is one that has not been well-characterized for its role as a dietary supplement. Therefore, the major objective of this experiment was to investigate two forms of valerate glycerides on diarrhea, intestinal physiology, and systemic immunity of weaned pigs experimentally infected with ETEC F18. Dietary treatments included a control diet and three additional diets supplemented with 0.075% monovalerin, 0.1% monovalerin, or 0.1% trivalerin, respectively. Piglets were weaned (21-24 d of age), individually housed, and experimental diets were fed throught the 28-day trial period. After a seven-day period, all piglets were inoculated on three consecutive days with 1010 CFU ETEC F18/3mL. Growth performance was monitored throughout the trial and daily diarrhea scores were recorded. Rectal swabs were collected for bacterial culture to confirm the presence or absence of β-hemolytic coliforms throughout the trial. Serum samples were collected and analyzed for inflammatory biomarkers on d 0, 3, 6, and 21 post-inoculation (PI) and untargeted metabolomics on d 6 PI. Intestinal mucosa and tissue sections were harvested from pigs sacrificed on d 7 PI for gene expression and histology analysis. All data, except for frequency of diarrhea and metabolomics, were analyzed by ANOVA using the PROC MIXED of SAS. Dietary trivalerin reduced (P < 0.05) the frequency of severe diarrhea over the entire trial period and the frequency of β-hemolytic coliforms on d 7 PI compared with control. The intestinal villus height on d 7 PI in jejunum tissue was increased (P < 0.05) in pigs fed trivalerin. The mRNA expression of TNF-α was decreased (P < 0.05) in the trivalerin group, while that of ZO1 was increased (P < 0.05) compared with control. Throughout the trial, serum TNF-α was reduced in pigs fed trivalerin compared with control. Serum metabolites, adenosine, inosine, and shikimic acid were reduced (P < 0.05) on d 6 PI in all treatment groups compared with control. In conclusion, the present results indicate supplementing dietary valerate glycerides exhibited beneficial impacts on diarrhea, inflammation, and intestinal gene expression of piglets during the post-weaning period.

Jianpi-Huatan-Huoxue-Anshen formula ameliorates gastrointestinal inflammation and microecological imbalance in chemotherapy-treated mice transplanted with H22 hepatocellular carcinoma.

Jianpi-Huatan-Huoxue-Anshen formula [Tzu-Chi cancer-antagonizing & life-protecting II decoction (TCCL)] is a Chinese medical formula that has been clinically shown to reduce the gastrointestinal side effects of chemotherapy in cancer patients and improve their quality of life. However, its effect and mechanism on the intestinal microecology after chemotherapy are not yet clear. To discover the potential mechanisms of TCCL on gastrointestinal inflammation and microecological imbalance in chemotherapy-treated mice transplanted with hepatocellular carcinoma (HCC). Ninety-six mice were inoculated subcutaneously with HCC cells. One week later, the mice received a large dose of 5-fluorouracil by intraperitoneal injection to establish a HCC chemotherapy model. Thirty-six mice were randomly selected before administration, and feces, ileal tissue, and ileal contents were collected from each mouse. The remaining mice were randomized into normal saline, continuous chemotherapy, Yangzheng Xiaoji capsules-treated, and three TCCL-treated groups. After treatment, feces, tumors, liver, spleen, thymus, stomach, jejunum, ileum, and colon tissues, and ileal contents were collected. Morphological changes, serum levels of IL-1β, IL-6, IL-8, IL-10, IL-22, TNF-α, and TGF-β, intestinal SIgA, and protein and mRNA expression of ZO-1, NF-κB, Occludin, MUC-2, Claudin-1, and IκB-α in colon tissues were documented. The effect of TCCL on the abundance and diversity of intestinal flora was analyzed using 16S rDNA sequencing. TCCL treatment improved thymus and spleen weight, thymus and spleen indexes, and body weight, decreased tumor volumes and tumor tissue cell density, and alleviated injury to gastric, ileal, and colonic mucosal tissues. Among proteins and genes associated with inflammation, IL-10, TGF-β, SIgA, ZO-1, MUC-2, and Occludin were upregulated, whereas NF-κB, IL-1β, IL-6, TNF-α, IL-22, IL-8, and IκB-α were downregulated. Additionally, TCCL increased the proportions of fecal Actinobacteria, AF12, Adlercreutzia, Clostridium, Coriobacteriaceae, and Paraprevotella in the intermediate stage of treatment, decreased the proportions of Mucipirillum, Odoribacter, RF32, YS2, and Rikenellaceae but increased the proportions of p_Deferribacteres and Lactobacillus at the end of treatment. Studies on ileal mucosal microbiota showed similar findings. Moreover, TCCL improved community richness, evenness, and the diversity of fecal and ileal mucosal flora. TCCL relieves pathological changes in tumor tissue and chemotherapy-induced gastrointestinal injury, potentially by reducing the release of pro-inflammatory factors to repair the gastrointestinal mucosa, enhancing intestinal barrier function, and maintaining gastrointestinal microecological balance. Hence, TCCL is a very effective adjuvant to chemotherapy.

Selenium nanoparticles affect chicken offspring's intestinal health better than other selenium sources

This study aimed to assess the effects of maternal diets containing various selenium (Se) sources on the intestinal mucosal function in the jejunum of chicken offspring. A total of 630, 18-wk-old Hy-Line Grey hens and 70 18-wk-old Hy-Line Grey breeders were randomly allocated into 7 groups, with 5 replicates in each group (18 hens and tow roosters). After 4 wk of Se depletion, the birds were fed either a nonsupplemented basal diet (control) or the same basal diet supplemented with 0.15 mg/kg selenium nanoparticles (Nano-Se), 0.30 mg/kg Nano-Se, 0.30 mg/kg selenocysteine (Sec), 0.30 mg/kg sodium selenite (SS), 0.30 mg/kg selenomethionine (SeMet), or 0.15 mg/kg Nano-Se + 0.15 mg/kg Sec, for 8 wk. Frtilized eggs were collected and incubated during the final week of the experiment. Jejunal tissues from embryonic d 18 and the hatch day were collected for analysis, and the 7-d survival rate of the offspring was recorded. Compared to the control, the maternal diet of 0.30 mg/kg Nano-Se, 0.30 mg/kg Sec, and 0.30 mg/kg SeMet significantly increased the survival of 7-day-old offspring (P < 0.05). The maternal diet supplemented with 0.30 mg/kg Nano-Se significantly increased intestinal villus height and the villus height/crypt depth ratio in chicks at embryonic d 18 and in 1-day-old (P < 0.05). The maternal diet containing 0.30 mg/kg Nano-Se and Sec increased the mRNA expression levels of tight junction proteins in 1-day-old offspring (P < 0.05). Supplemental 0.30 mg/kg Nano-Se significantly increased the mRNA expression levels of marker genes in intestinal enteroendocrine, stem, and Paneth cells (P < 0.05). In 1-day-old chicks, the number of intestinal goblet cells, as well as the mRNA expression levels of intestinal mucin2 (Muc2) and goblet cell differentiation factors (Spdef and C-myc), were the highest in diets supplemented with 0.30 mg/kg Nano-Se. Moreover, the expression levels of intestinal Muc2 and Spdef in chicks at embryonic d 18 was the highest with 0.30 mg/kg Nano-Se supplementation (P < 0.05). Supplementing with 0.30 mg/kg Nano-Se significantly reduced reactive oxygen species levels and decreased the mRNA expression levels of apoptosis-related genes in 1-day-old chicks (P < 0.05). Additionally, 0.30 mg/kg Nano-Se supplementation significantly down-regulated NLRP3 pathway gene expression in 1-day-old chicks (P < 0.05). In conclusion, maternal dietary supplementation with Nano-Se improved jejunal microarchitecture, antioxidant levels, and the expression of tight-junction protein in chicken offspring along with supporting goblet cell development by inhibiting the NLRP3 signaling pathway.

The interactivity of sources and dietary levels of resistant starches – impact on growth performance, starch, and nutrient digestibility, digesta oligosaccharides profile, cecal microbial metabolites, and indicators of gut health in broiler chickens

In a 21-d study, 480 Cobb 500 (off-sex) male broiler chicks were used to investigate the effects of feeding different sources and levels of resistant starches (RS) on growth performance, nutrient and energy utilization, and intestinal health in broiler chickens. The birds were allocated to 10 dietary treatments in a 3 × 3 + 1 factorial arrangement. The factors were 3 RS-sources (RSS): banana starch (BS), raw potato starch (RPS), and high-amylose corn starch (HCS); each at 3 levels (RSL) 25, 50, or 100 g/kg plus a corn-soybean meal control diet. Birds and feed were weighed on d 0, 8, and 21. On d 21, samples of jejunal tissue and digesta were collected for chemical analysis. Data were analyzed using the mixed model procedure of JMP with factor levels nested with the control. In the 0 to 21 phase, the birds fed the RPS diets had higher (P = 0.011) FI than those fed HCS or control diets, and FCR was greater (P = 0.030) in birds that received BS diets than in other diets. RSS × RSL was significant (P < 0.05) for total tract nutrient retention, AME, and AMEn on d 21. The starch digestibility was higher (P < 0.001) in birds that received the control diet than in RS diets, and decreased as RS levels increased, except for HCS. The apparent metabolizable energy (AME) and nitrogen-corrected AME (AMEn) were higher (P < 0.001) in birds fed 100 g/kg HCS diet, with both decreasing with increasing levels of BS and RPS, except for HCS. Relative ileal oligosaccharides profile showed significant (P < 0.05) RSS × RSL with a higher relative abundance of Hex(3) (P = 0.01) and Pent(3) (P = 0.001) in HCS diets. In conclusion, RS may influence gut health and growth performance in broiler chickens through modulation of cecal SCFA and nutrient digestion, but these depend largely on the botanical origin and concentrations of individual RS.

Bacillus amyloliquefaciens TL promotes gut health of broilers by the contribution of bacterial extracellular polysaccharides through its anti-inflammatory potential.

The focal point of probiotic efficacy and a crucial factor influencing poultry cultivation lies in the level of intestinal inflammation. In conventional farming processes, the reduction of intestinal inflammation generally proves advantageous for poultry growth. This study investigated the impact of Bacillus amyloliquefaciens TL (B.A.-TL) on inflammatory factor expression at both tissue and cellular levels, alongside an exploration of main active secondary metabolites. The results demonstrated that broiler feeding with a basal diet containing 4 × 109 CFU/kg B.A.-TL markedly enhanced chicken growth performance, concomitant with a significant decrease in the expression of genes encoding inflammatory cytokines (e.g., CCL4, CCR5, XCL1, IL-1β, IL-6, IL-8, LITAF, and LYZ) in jejunum and ileum tissues. The extracellular polysaccharides of B.A.-TL (EPS-TL) exhibited notable suppression of elevated inflammatory cytokine expression induced by Escherichia coli O55 lipopolysaccharides (LPS) in chicken macrophage-like cells (HD11) and primary chicken embryonic small intestinal epithelial cells (PCIECs). Moreover, EPS-TL demonstrated inhibitory effect on NF-κB signaling pathway activation. These findings suggested that the metabolic product of B.A.-TL (i.e., EPS-TL) could partly mitigate the enhanced expression of inflammatory factors induced by LPS stimulation, indicating its potential as a key component contributing to the anti-inflammatory effects of B.A.-TL.

PSI-28 Effect of supplementing a Bacillus spp.-based probiotic on intestinal tight junction proteins expression and abundance of selected microorganisms in heat stressed pigs

Abstract The maintenance of small intestine epithelial barrier function, depending in part on tight junction proteins (TJP), prevents the system from invasion of undesired materials but may become altered in heat stressed pigs. Heat stress (HS) also appears to modify the composition of intestinal microbiota that, combined with a weak barrier, may cause health deterioration of pigs. Probiotics, in contrast, can help animals to maintain a healthy intestinal environment. A study, conducted with pigs [n = 80; body weight (BW) = 21.6 ± 3.4 kg], analyzed the effect of HS and the supplementation of a Bacillus spp-based probiotic on gene expression of the TJP occludin, claudins, and TJP1 in jejunum and ileum. Abundance of Lactobacillus sp., Bifidobacterium sp., Bacillus subtilis and E. coli in ileal content was also analyzed. Treatments were thermal neutral (TN) pigs fed a control diet without probiotic (TN-C) or with 0.05% probiotic (TN-P), and HS pigs fed a control without probiotic (HS-C) or with 0.05% probiotic (HS-P). The control diet was based on wheat and soybean meal, added with free Lys, Thr, Met, and Val. Feed and water were freely available during the 21-d study. At termination, six pigs per treatment were sacrificed and samples from jejunum and ileum tissue were collected to analyze TJP gene expression; ileal content was collected to analyze abundance of intestinal microorganisms. Ambient temperature inside TN and HS rooms ranged from 19 to 25°C and 30 to 38.5°C, respectively. In jejunum, claudin2 expression was greater in TN-P than in TN-C (P = 0.05) but there was no effect of HS. In ileum, HS increased claudin3 but decreased occludin gene expression (P &amp;lt; 0.05). HS did not affect TJP1gene expression (P &amp;gt; 0.10). Probiotic supplementation did not affect gene expression of TJP (P &amp;gt; 0.10). Relative to TN-C pigs, abundance of Lactobacillus sp. and Bifidobacterium sp. tended to decrease in HS-C pigs (P &amp;lt; 0.10), and that of Bifidobacterium sp. decreased in HS-P pigs (P &amp;lt; 0.05). Abundance of E. coli tended to be greater (P &amp;lt; 0.10) in both HS-C and HS-P pigs, than in the TN-C pigs. Abundance of Bacillus sp. was greater (P &amp;lt; 0.01) in HS pigs fed the diet supplemented with the probiotic, compared with the TN-C (P &amp;lt; 0.01) and HS-C (P &amp;lt; 0.05) pigs. In conclusion, these data indicate that HS affects TJP gene expression and composition of intestinal microbiota, but supplementing 0.05% of a probiotic based on Bacillus spp. did not affect the abundance of measured microbiota except for Bacillus sp. under HS conditions.

500 Effect of different iron sources on growth performance, gut health, and microbiota in nursery pigs

Abstract This study evaluated the effect of different iron sources, including iron polysaccharide complexes (SQM iron) and ferrous sulfate (FS) on growth performance, gut health, pathogenic bacteria load, colon fecal microbiota, and fecal iron concentration in nursery pigs. Nursery pigs (n = 320, TN70 cross, Topigs Norsvin Canada) weaned at 21 ± 2 d were randomly assigned to four treatments arranged in 2 x 2 factorial, with the factors being sanitation conditions (clean vs. dirty) and iron sources (SQM iron vs. FS, each at 100 mg iron/kg) to give 8 pens (10 piglets per pen) per treatment. Pigs were fed their respective diets in two phases over a 28-d period. Fecal scores, body weight (BW) and feed intake were recorded weekly. Iron content, gene expression of butyryl-CoA:acetate CoA-transferase and bisulfite reductase, and pathogenic bacteria counts were determined in fresh fecal samples collected on d 0, d 14, and d 21. On d 28, one pig randomly selected from each pen was euthanized to sample colon digesta for microbiota analysis and jejunal tissue and serum for gut integrity and permeability analysis. Data were analyzed using PROC MIXED of SAS followed by Tukey’s multiple comparison test except for raw 16s sRNA sequences, which were processed using QIIME2 and analyzed with Kruskal-Wallis test. Overall, from d 0 to14 SQM iron resulted in higher average daily gain (ADG) and average daily feed intake (ADFI; P &amp;lt; 0.05). SQM iron resulted in higher fecal iron at d 21 and improved fecal scores at d 27 (P &amp;lt; 0.05). SQM iron increased the relative abundance of fecal Campylobacter jejuni when comparing d 21 to d 0, and O157 E. Coli when comparing d 14 to d 0 (P &amp;lt; 0.05). Ferrous sulfate resulted in greater changes in relative abundance of butyryl-CoA:acetate CoA-transferase and bisulfate reductase than the SQM iron treatment under clean conditions when comparing d 21 to d 0 (P &amp;lt; 0.05). Dietary iron treatments did not significantly affect gastrointestinal integrity and permeability in jejunum and did not alter fecal microbiota composition on d 28. In conclusion, substituting 100 mg/kg added iron as FS with equal concentration of SQM iron improved ADG in the initial 2 wk and overall ADFI and promoted gut health indicators.

Alginate Oligosaccharides Enhance Gut Microbiota and Intestinal Barrier Function, Alleviating Host Damage Induced by Deoxynivalenol in Mice

BackgroundAlginate oligosaccharides (AOS) exhibits notable effects in terms of anti-inflammatory, antibacterial, and antioxidant properties. Deoxynivalenol (DON) has the potential to trigger intestinal inflammation by upregulating pro-inflammatory cytokines and apoptosis, thereby compromising the integrity of the intestinal barrier function and perturbing the balance of the gut microbiota. ObjectivesWe assessed the impact of AOS on mitigating DON-induced intestinal damage and systemic inflammation in mice. MethodsAfter a 1-wk acclimatization period, the mice were divided into 4 groups. For 3 wk, the AOS and AOS + DON groups were gavaged daily with 200 μL of AOS [200 mg/kg body weight (BW)], whereas the CON and DON groups received an equivalent volume of sterile Phosphate-Buffered Saline (PBS). Subsequently, for 1 wk, the DON and AOS + DON groups received 100 μL of DON (4.8 mg/kg BW) daily, whereas the control (CON) and AOS groups continued receiving PBS. ResultsAfter administering DON via gavage to mice, there was a significant decrease (P < 0.05) in body weights compared with the CON group. Interestingly, AOS exhibited a tendency to mitigate this weight loss in the AOS + DON group. In the feces of mice treated with both AOS and DON, the concentration of DON significantly increased (P < 0.05) compared with the DON group alone. Histological analysis revealed that DON exposure caused increased intestinal damage, including shortened villi and eroded epithelial cells, which was ameliorated by presupplementation with AOS, alleviating harm to the intestinal barrier function. In both jejunum and colon tissues, DON exposure significantly reduced (P < 0.05) the expression of tight junction proteins (claudin and occludin in the colon) and the mucin protein mucin 2, compared with the CON group. Prophylactic administration of AOS alleviated these reductions, thereby improving the expression levels of these key proteins. Additionally, AOS supplementation protected DON-exposed mice by increasing the abundance of probiotics such as Bifidobacterium, Faecalibaculum, and Romboutsia. These gut microbes are known to enhance (P < 0.05) anti-inflammatory responses and the production of short-chain fatty acids (SCFAs), including total SCFAs, acetate, and valerate, compared with the DON group. ConclusionsThis study unveils that AOS not only enhances gut microbiota and intestinal barrier function but also significantly mitigates DON-induced intestinal damage.

Identification of COL3A1 as a candidate protein involved in the crosstalk between obesity and diarrhea using quantitative proteomics and machine learning

BackgroundIncreasing epidemiologic studies have shown a positive correlation between obesity and chronic diarrhea. Nevertheless, the precise etiology remains uncertain. MethodsWe performed a comprehensive proteomics analysis utilizing the data-independent acquisition (DIA) technique on jejunal tissues from patients with obesity and chronic diarrhea (OD, n = 33), obese patients (OB, n = 10), and healthy controls (n = 8). Differentially expressed proteins (DEPs) in OD vs. control and OD vs. OB comparisons were subjected to pathway enrichment and protein-protein interaction (PPI) network analysis. Machine learning algorithms were adopted on overlapping DEPs in both comparisons. The candidate protein was further validated using Western blot, immunohistochemistry (IHC), and in vitro experiments. ResultsWe identified 189 and 228 DEPs in OD vs. control and OD vs. OB comparisons, respectively. DEPs in both comparisons were co-enriched in extracellular matrix (ECM) organization. Downregulated DEPs were associated with tight junction and ECM-receptor interaction in OD vs. control and OD vs. OB comparisons, respectively. Machine learning algorithms selected 3 proteins from 14 overlapping DEPs in both comparisons, among which collagen alpha-1(III) chain (COL3A1) was identified as a core protein in PPI networks. Western blot and IHC verified the expression of COL3A1. Moreover, the tight junction-related proteins decreased after the knockdown of COL3A1 in Caco2 intestinal cells upon PA challenge, consistent with the proteomics results. ConclusionsWe generated in-depth profiling of a proteomic dataset from samples of OD patients and provided unique insights into disease pathogenesis. COL3A1 was involved in the crosstalk between obesity and intestinal homeostasis via the ECM-receptor interaction pathway.

Anticoccidial and Antioxidant Activities of an Ethanolic Extract of Teucrium polium Leaves on Eimeria papillate-Infected Mice.

Eimeria spp. are responsible for the economic loss of both domestic and wild animals due to coccidiosis, the most common parasitic disease. The resistance to currently available drugs used to treat coccidiosis has been proven. Medicinal plants that contain physiologically active phytochemicals have been widely used in traditional medicine. Teucrium polium leaf extract (TPLE) has been shown to exhibit pharmacological, antioxidant, and anticoccidial properties in different experiments. Here, our investigation focused on how T. polium leaf extract affected the way that Eimeria papillate caused intestinal injury in mice. Thirty-five male Swiss albino mice were divided into seven groups, as follows: group I: untreated and uninfected (negative control); group II: uninfected, treated group with TPLE (150 mg/kg b.w); and group III: infected untreated (positive control). Groups III-VII were orally administered 103 sporulated E. papillata oocysts. A total of 60 min after infection, groups IV-VI were treated for five successive days with 50, 150, and 250 mg/kg b.w TPLE, respectively, while group VII was treated with amprolium (120 mg/kg b.w.). The mice had been euthanized on the fifth day post-infection, and the jejunum tissues were prepared for histology and oxidative stress studies. A total of 150 mg/kg of TPLE was the most effective dosage, significantly decreasing oocyst output by about 80.5%, accompanied by a significant reduction in the number of developmental parasitic phases in jejunal sections. In addition, the decrease in the number of goblet cells in the jejuna of mice raised after treatment. Also, TPLE greatly diminished the body weight loss of infected mice. Moreover, our research proved that TPLE reduced oxidative damage due to E. papillata infection via decreasing intestinal malondialdehyde (MDA) and nitric oxide (NO) levels and increasing reduced superoxide dismutase (SOD) and glutathione (GSH) levels. These results demonstrated that TPLE had potent anticoccidial properties. TPE's efficacy as a natural antioxidant has also been demonstrated in reducing oxidative stress and enhancing antioxidant systems to mitigate biochemical and histological changes in the jejunum caused by E. papillata.

Stimbiotic and wheat bran inclusion in maize or wheat-based diets for broiler chickens; effects on jejunal histomorphology, digesta oligosaccharides and caecal short-chain fatty-acids profiles

ABSTRACT 1. A stimbiotic (STB) is any feed additive that stimulates caeca fibre fermentation, although the additive itself contributes little to the caeca short-chain fatty acid (SCFA) production. A 42 d experiment investigated the interactive effects of STB and wheat bran (WB) in broiler chickens receiving maize or wheat-based diets. 2. The treatments were arranged in a 2 × 2 × 2 factorial (eight replicates each), the dietary factors being diet (maize-SBM or wheat-SBM), STB (with or without) and WB (0 or 50 g/kg). Jejunal tissue, gizzard, jejunal and ileal digesta and caecal contents were collected on d 18 and 42. 3. Gizzard pH tended to decrease with STB (p = 0.06) supplementation and was lower in birds fed wheat- compared to maize-based diets on d 18 (p < 0.05). Birds receiving diets with WB had higher jejunum pH on d 18 (p < 0.05). 4. Total short-chain fatty acids (SCFA) in the caeca on d 18 and isobutyrate on d 42 were higher (p < 0.05) for maize compared with wheat-based diets. However, on d 42, acetate, butyrate and total SCFA were higher (p < 0.05) for wheat-based compared with maize-based diets. 5. On d 18, STB and WB inclusion increased villi height (VH; p < 0.05) and VH to crypt depth ratio (VH/CD), respectively (p < 0.05). On d 42, VH (p < 0.05) and VH/CD were higher in wheat-based diets (p < 0.05). The VH/CD ratio was lower with STB supplementation (p < 0.05). Marker-corrected pentose oligosaccharides (Pent)4 and (Pent)5 concentrations in the ileal digesta were reduced (p < 0.05) with STB supplementation. In addition, STB decreased (Pent)3 concentration in maize-, but not wheat-based diets (p < 0.05). 6. In conclusion, both WB and STB influenced gastrointestinal pH and jejunum histomorphology of broilers without increasing oligosaccharide concentration in the ileum and SCFA in the caeca.

Cell-Free Supernatant of Bacillus subtilis G2B9-Q Improves Intestinal Health and Modulates Immune Response to Promote Mouse Recovery in Clostridium perfringens Infection.

Clostridium perfringens is one of the critical causative agents causing diarrhea in piglets, with significant economic losses to the pig industry. Under normal gut microbiota homeostasis and well-managed barns, diarrhea caused by C. perfringens could be controlled. Some reports show that probiotics, such as Bacillus subtilis, are beneficial in preventing necrotic enteritis (NE) in chickens, but few reports on piglets. Clostridium perfringens was found in the piglets' diarrhea with intestinal microbiota dysbiosis in our survey. Bacillus subtilis G2B9-Q, which was isolated from the feces of healthy pigs, was found to have anti-Clostridium activity after screening. Clostridium perfringens was used to challenge mice by intraperitoneal injection for modeling to evaluate the anti-infective activity of cell-free supernatant (CFS) of B. subtilis G2B9-Q and different concentrations of B. subtilis G2B9-Q by oral administration. The results showed that G2B9-Q can mitigate intestinal lesions caused by C. perfringens infection, reduce inflammatory reactions, and modulate intestinal microbiota. The CFS of G2B9-Q can alleviate the pathological damage of intestinal tissues caused by C. perfringens infection, reduce the concentration of TNF-α and IL-10 in the sera of mice, as well as the relative expression levels of alpha toxin (CPA), perfringolysin O (PFO) toxin, IL-10, IL-22, andTNF-α in the jejunum and colon tissues, and alleviate the changes in gut microbiota structure caused by C. perfringens infection, which showed better therapeutic effects and indicated that the metabolites of G2B9-Q are essential mediators for their beneficial effects. Therefore, the CFS of G2B9-Q could potentially replace antibiotics in treating C. perfringens infection.