Intestinal Development Research Articles

PSIII-15 Effect of dietary fatty acids composition on growth performance, fecal consistency, and plasma fatty acid profile of weaned pigs

Abstract Dietary lipids are an important source of energy and fatty acids (FA), which have critical roles in mucosal immune responses, epithelial barrier functions, and overall intestinal development. Therefore, including fat in nursery pig diets can be used as a nutritional strategy to optimize pig growth performance and intestinal health after weaning. This study evaluated the effects of two different dietary fatty acid compositions derived from conventional soybean oil (CSO) and/or high oleic soybean oil (HOSO) supplementation on growth performance, fecal consistency, and plasma FA profile of weaned pigs. A total of 208 weaned pigs (body weights = 4.96 ± 0.8 kg) were used in a randomized complete block design and allotted to 16 pens of 13 pigs each. Experimental diets met NRC (2012) requirements and were provided in a 3-phase nursery feeding program: Phase 1 (d 0-d7), Phase 2 (d 8-d21), and Phase 3 (d 22-d42). Pens were assigned to one of two dietary treatments consisting of two dietary FA compositions derived from the supplementation of 3% of either CSO (20% Oleic acid; 50% Linoleic acid; 6% Linolenic acid) or HOSO (44% Oleic acid; 30% Linoleic acid; 3% Linolenic acid). Feed disappearance and body weight were measured at d 0, 21, and 42. Pen fecal score was assigned daily from d 0 to 21 and then 3 times per week until d 42. Blood samples were collected on d 21 (2 pigs/pen) for plasma FA profile analysis. Performance data were analyzed using T-TEST and fecal score using PROC FREQ procedures of SAS. From d 0 to 21, pigs fed CSO supplemented diets had a greater (P ≤ 0.05) body weight (8.6 vs. 7.4 kg) and daily gain (0.17 vs. 0.12 kg). In addition, daily feed intake and feed efficiency tended to be 23% and 16% greater in CSO-fed pigs, respectively. From d 21 to d 42, there were no differences between dietary treatments for growth performance. Overall nursery period, pigs fed CSO had greater (P ≤ 0.1) daily gain than HOSO pigs (0.34 vs. 0.30 kg). Pigs fed CSO had a greater (P ≤ 0.05) plasma concentration of palmitic, linoleic, and linolenic FA while pigs fed HOSO had a greater (P ≤ 0.05) plasma concentration of palmitoleic and oleic FA. There were no differences in fecal scores in wk 1 and 2. However, pigs fed CSO had less incidence of soft and watery feces (χ2 < 0.1) in wk 3 to 6. In conclusion, a greater dietary content of linoleic and linolenic FA promoted pig growth performance and reduced incidence of diarrhea after weaning compared with greater content of oleic FA.

PSIV-A-7 Effects of late gestational nutrient restriction of beef heifers on small intestinal crypt depth in neonatal calves

Abstract We hypothesized that late gestational nutrient restriction would impair small intestinal development in neonatal calves, including crypt cell depth and proliferation. Fall-calving crossbred beef heifers [body weight (BW) = 466 ± 30 (SD) kg; BCS: 5.4 ± 0.5] bred to a single sire were individually-fed 100% (control; n = 10) or 70% (nutrient restricted; n = 12) of NASEM metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth from d 160 of gestation to calving. Calves were removed from their dams immediately post-calving and fed artificial colostrum and milk replacer relative to body weight until euthanasia at 48.7 ± 0.4 h of age. The gastrointestinal tract was removed, and proximal jejunum samples were dissected from each calf and immersion-fixed in 10% neutral buffered formalin. Tissues were embedded in paraffin blocks and cut into 5-µm sections that were mounted on glass slides. Slides were incubated with Ki-67 (marker of nuclear proliferation) and counterstained with Hematoxylin using an autostainer. Images were taken at 5× magnification using an upright microscope. QuPath software was used to measure crypt depth, starting at the base of the crypt region and ending at the furthest cluster of Ki-67 positive cells, perpendicular to the base of the mucosa. For each calf, 10 crypt depth measurements were taken from 4 images, resulting in 40 crypt depth measurements that were averaged. Nutritional plane was a fixed effect in the model, and maternal sire, date of study initiation, and calf sex (when P < 0.25) were considered covariates. We previously reported that calves born to nutrient restricted dams weighed less (P ≤ 0.05) at birth and 48 h than calves born to control dams. Small intestinal mass tended to be less (P = 0.11) in offspring of nutrient restricted dams but was not affected (P = 0.58) when expressed relative to body weight. Calves born to nutrient restricted dams also tended to have less (P = 0.11) small intestinal mass relative to brain weight. Maternal nutrient restriction did not affect (P = 0.95) jejunal crypt depth. Additionally, crypt depth relative to body weight was not affected (P = 0.20) by nutrient restriction. Although crypt depth was not affected, further analysis of proliferation and morphology will help to understand the relationship between maternal nutrient restriction and small intestine development in neonatal calves.

Sorbitol Destroyed Intestinal Microfold Cells (M Cells) Development through Inhibition of PDE4-Mediated RANKL Expression.

Microfold cells (M cells) are specific intestinal epithelial cells for monitoring and transcytosis of antigens, microorganisms, and pathogens in the intestine. However, the mechanism for M-cell development remained elusive. Real-time polymerase chain reaction, immunofluorescence, and western blotting were performed to analyze the effect of sorbitol-regulated M-cell differentiation in vivo and in vitro, and luciferase and chromatin Immunoprecipitation were used to reveal the mechanism through which sorbitol-modulated M-cell differentiation. Herein, in comparison to the mannitol group (control group), we found that intestinal M-cell development was inhibited in response to sorbitol treatment as evidenced by impaired enteroids accompanying with decreased early differentiation marker Annexin 5, Marcksl1, Spib, sox8, and mature M-cell marker glycoprotein 2 expression, which was attributed to downregulation of receptor activator of nuclear factor kappa-В ligand (RANKL) expression in vivo and in vitro. Mechanically, in the M-cell model, sorbitol stimulation caused a significant upregulation of phosphodiesterase 4 (PDE4) phosphorylation, leading to decreased protein kinase A (PKA)/cAMP-response element binding protein (CREB) activation, which further resulted in CREB retention in cytosolic and attenuated CREB binds to RANKL promoter to inhibit RANKL expression. Interestingly, endogenous PKA interacted with CREB, and this interaction was destroyed by sorbitol stimulation. Most importantly, inhibition of PDE4 by dipyridamole could rescue the inhibitory effect of sorbitol on intestinal enteroids and M-cell differentiation and mature in vivo and in vitro. These findings suggested that sorbitol suppressed intestinal enteroids and M-cell differentiation and matured through PDE4-mediated RANKL expression; targeting to inhibit PDE4 was sufficient to induce M-cell development.

Effects of feed conditioning temperature on pellet quality, growth performance, intestinal development and blood parameters of geese from 1 to 28 days of age

Effects of feed conditioning temperature on pellet quality, growth performance, intestinal development and blood parameters of geese from 1 to 28 days of age

Impact of the thyroid hormone T3 and its nuclear receptor TRα1 on colon cancer stem cell phenotypes and response to chemotherapies

Colorectal cancers (CRCs) are highly heterogeneous and show a hierarchical organization, with cancer stem cells (CSCs) responsible for tumor development, maintenance, and drug resistance. Our previous studies showed the importance of thyroid hormone-dependent signaling on intestinal tumor development and progression through action on stem cells. These results have a translational value, given that the thyroid hormone nuclear receptor TRα1 is upregulated in human CRCs, including in the molecular subtypes associated with CSC features. We used an established spheroid model generated from the human colon adenocarcinoma cell line Caco2 to study the effects of T3 and TRα1 on spheroid formation, growth, and response to conventional chemotherapies. Our results show that T3 treatment and/or increased TRα1 expression in spheroids impaired the response to FOLFIRI and conferred a survival advantage. This was achieved by stimulating drug detoxification pathways and increasing ALDH1A1-expressing cells, including CSCs, within spheroids. These results suggest that clinical evaluation of the thyroid axis and assessing TRα1 levels in CRCs could help to select optimal therapeutic regimens for patients with CRC.Proposed mechanism of action of T3/TRα1 in colon cancer spheroids. In the control condition, TRα1 participates in maintaining homeostatic cell conditions. The presence of T3 in the culture medium activates TRα1 action on target genes, including the drug efflux pumps ABCG2 and ABCB1. In the case of chemotherapy FOLFIRI, the increased expression of ABC transcripts and proteins induced by T3 treatment is responsible for the augmented efflux of 5-FU and Irinotecan from the cancer cells. Taken together, these mechanisms contribute to the decreased efficacy of the chemotherapy and allow cells to escape the treatment. Created with BioRender.com.

Gelsenicine disrupted the intestinal barrier of Caenorhabditis elegans

Gelsemium elegans Benth. (G. elegans) is a traditional medicinal herb that has anti-inflammatory, analgesic, sedative, and detumescence effects. However, it can also cause intestinal side effects such as abdominal pain and diarrhea. The toxicological mechanisms of gelsenicine are still unclear. The objective of this study was to assess enterotoxicity induced by gelsenicine in the nematodes Caenorhabditis elegans (C. elegans). The nematodes were treated with gelsenicine, and subsequently their growth, development, and locomotion behavior were evaluated. The targets of gelsenicine were predicted using PharmMapper. mRNA-seq was performed to verify the predicted targets. Intestinal permeability, ROS generation, and lipofuscin accumulation were measured. Additionally, the fluorescence intensities of GFP-labeled proteins involved in oxidative stress and unfolded protein response in endoplasmic reticulum (UPRER) were quantified. As a result, the treatment of gelsenicine resulted in the inhibition of nematode lifespan, as well as reductions in body length, width, and locomotion behavior. A total of 221 targets were predicted by PharmMapper, and 731 differentially expressed genes were screened out by mRNA-seq. GO and KEGG enrichment analysis revealed involvement in redox process and transmembrane transport. The permeability assay showed leakage of blue dye from the intestinal lumen into the body cavity. Abnormal mRNAs expression of gem-4, hmp-1, fil-2, and pho-1, which regulated intestinal development, absorption and catabolism, transmembrane transport, and apical junctions, was observed. Intestinal lipofuscin and ROS were increased, while sod-2 and isp-1 expressions were decreased. Multiple proteins in SKN-1/DAF-16 pathway were found to bind stably with gelsenicine in a predictive model. There was an up-regulation in the expression of SKN-1:GFP, while the nuclear translocation of DAF-16:GFP exhibited abnormality. The UPRER biomarker HSP-4:GFP was down-regulated. In conclusion, the treatment of gelsenicine resulted in the increase of nematode intestinal permeability. The toxicological mechanisms underlying this effect involved the disruption of intestinal barrier integrity, an imbalance between oxidative and antioxidant processes mediated by the SKN-1/DAF-16 pathway, and abnormal unfolded protein reaction.

Antibiotic Cocktail Effects on Intestinal Microbial Community, Barrier Function, and Immune Function in Early Broiler Chickens

This study investigated the effects of an antibiotic cocktail on intestinal microbial composition, mechanical barrier structure, and immune functions in early broilers. One-day-old healthy male broiler chicks were treated with a broad-spectrum antibiotic cocktail (ABX; neomycin, ampicillin, metronidazole, vancomycin, and kanamycin, 0.5 g/L each) or not in drinking water for 7 and 14 days, respectively. Sequencing of 16S rRNA revealed that ABX treatment significantly reduced relative Firmicutes, unclassified Lachnospiraceae, unclassified Oscillospiraceae, Ruminococcus torques, and unclassified Ruminococcaceae abundance in the cecum and relative Firmicutes, Lactobacillus and Baccillus abundance in the ileum, but significantly increased richness (Chao and ACE indices) and relative Enterococcus abundance in the ileum and cecum along with relatively enriched Bacteroidetes, Proteobacteria, Cyanobacteria, and Enterococcus levels in the ileum following ABX treatment for 14 days. ABX treatment for 14 days also significantly decreased intestinal weight and length, along with villus height (VH) and crypt depth (CD) of the small intestine, and remarkably increased serum LPS, TNF-α, IFN-γ, and IgG levels, as well as intestinal mucosa DAO and MPO activity. Moreover, prolonged use of ABX significantly downregulated occludin, ZO-1, and mucin 2 gene expression, along with goblet cell numbers in the ileum. Additionally, chickens given ABX for 14 days had lower acetic acid, butyric acid, and isobutyric acid content in the cecum than the chickens treated with ABX for 7 days and untreated chickens. Spearman correlation analysis found that those decreased potential beneficial bacteria were positively correlated with gut health-related indices, while those increased potential pathogenic strains were positively correlated with gut inflammation and gut injury-related parameters. Taken together, prolonged ABX application increased antibiotic-resistant species abundance, induced gut microbiota dysbiosis, delayed intestinal morphological development, disrupted intestinal barrier function, and perturbed immune response in early chickens. This study provides a reliable lower-bacteria chicken model for further investigation of the function of certain beneficial bacteria in the gut by fecal microbiota transplantation into germ-free or antibiotic-treated chickens.

Exploring functional metabolites and proteomics biomarkers in late-preterm and natural-born pigs.

Pigs are often used to study the intestinal development of newborns, particularly as preterm pig models that mimic the intestinal growth of human preterm infants. Neonatology's study of delivery mode's impact on neonatal development is crucial. We established 14 newborn pigs delivered via cesarean sections (C-section, at 113 days of gestational age, CS group) and 8 naturally born pigs were used as controls (at 114 days of gestational age, NF group). The impact of two alternative delivery procedures (C-section and natural birth) on the levels of short-chain fatty acids (SCFAs) and organic acids in the hepatic and intestines of newborn pigs were compared using metabolomics. The underlying molecular pathways are examined at the "protein-metabolite" level by integrating proteomic data. The findings demonstrated that the mode of delivery changed the metabolism of SCFAs in newborn pigs, perhaps by affecting the physiology levels of cyclic intermediates such as lactate and malate in the pyruvate metabolic pathway. Additionally, by participating in the fatty acid metabolism pathway, two distinct proteins (FASN and HSD17B4) may impact the physiological concentration of these tiny metabolites. In conclusion, this study provided reliable animal model data for understanding the physiological SCFA metabolic information and its affecting mechanism of large-gestational age preterm infants.

The Roles of Polyamines in Intestinal Development and Function in Piglets.

The gastrointestinal tract plays crucial roles in the digestion and absorption of nutrients, as well as in maintenance of a functional barrier. The development and maturation of the intestine is important for piglets to maintain optimal growth and health. Polyamines are necessary for the proliferation and growth of enterocytes, which play a key role in differentiation, migration, remodeling and integrity of the intestinal mucosa after injury. This review elaborates the development of the structure and function of the intestine of piglets during embryonic, suckling and weaning periods, the utilization and metabolism of polyamines in the intestine, as well as the role of polyamines in intestinal development and mucosal repair. The nutritional intervention to improve intestinal development and functions by modulating polyamine metabolism in piglets is also put forward. These results may help to promote the adaption to weaning in pigs and provide useful information for the development and health of piglets.

Selection of a multi-species starter culture for mustard seed fermentation to enhance polyunsaturated fatty acids and improve gastrointestinal health markers

Fermented mustard seed is a unique delicacy from Assam, India, with many health benefits. The fermentation process involves a complex interplay of various microbes that give the final product its desired characteristics. A comprehensive understanding of fermented mustard's microbial and metabolite composition can aid in optimizing the fermentation process, enabling the retention of maximum beneficial effects. This study reports the traditional fermentation process of mustard seeds, the composition of its native microbe-metabolite, and the development of a starter culture that increases the polyunsaturated fatty acid (PUFA) content. The fermented mustard seeds have a complex bacterial ecosystem comprising of Staphylococcus, Bacillus, Mammaliicoccus, Weisella, Corynebacterium, Aerococcus, Ruoffia, Paenibacillus, and Enterococcus. Moreover, untargeted metabolomics reveals that it is a rich source of functional metabolites, especially PUFA. Based on multi-omics approaches, the study deduced the core bacterial composition to be S. succinus, S. saprophyticus, and B. subtilis, with a strong positive correlation (p < 0.05) with the synthesis of PUFA. Optimized fermentation experiments of mustard seed with the core bacterial group resulted in an enhanced PUFA content of 52.9%. Transcriptome study on HT-29 cells treated with optimized fermented mustard seed extract exhibited differential expression of genes linked to cellular pathways associated with gastrointestinal health, such as gut homeostasis, intestinal barrier integrity, immuno-modulatory, and neuronal development. This study revealed an omics-guided strategy for the development of starter-assisted controlled fermentation with experimental validation on fermented mustard seed.

Improving effect of N-acetylcysteine on growth and intestinal health in juvenile Cyprinus carpio involve the Nrf2/MAPK/NF-κB pathways

N-acetylcysteine (NAC) plays a beneficial role in improving animal growth and health. The current study evaluated the impact of dietary supplementation with NAC on the growth performance, physiological status and intestinal health of juvenile Cyprinus carpio. The experiment involved 5 groups, with 3 replicates in each group, and each replicate group containing 30 juvenile Cyprinus carpio (average weight 2.06 ± 0.01 g). Five experimental diets (36% crude protein and 9% crude lipid) were prepared by supplementing graded levels (0, 0.15%, 0.30%, 0.45% and 0.60%) of NAC in the basal diet. Subsequently, these 5 diets were utilized for an 56 d feeding trial with Cyprinus carpio. During the feeding trial, the water temperature was maintained between 24 and 28 °C, and the feeding rate was 3%–5% of body weight. After the feeding trial, fish were fasted for 24 h, anesthetized, and dissected on ice trays to collect samples for subsequent analysis. Results showed that NAC (0.30%, 0.45% and 0.60% diets) significantly improved the growth performance and intestinal mucosa development in Cyprinus carpio, and inhibited apoptosis. Similarly, NAC (0.30%, 0.45% and 0.60% diets) could enhance the homeostasis of the redox defense system in intestine of Cyprinus carpio, which is linked to enhanced glutathione metabolism, scavenging of reactive oxygen species and activation of Nrf2 pathway. Furthermore, dietary NAC improved anti-inflammatory and immunomodulatory functions, and involved in the regulation of MAPK/NF-κB signaling pathway. Summary, NAC (0.30%, 0.45% and 0.60% diets) could significantly promote the growth and intestinal health in Cyprinus carpio. These findings provide partial available information for promoting the utilization of NAC in aquafeed and improving the ecological and healthy cultivation of Cyprinus carpio.

A Preliminary Study on the Effect of Replacing Part of Fishmeal by Brewer’s Yeast in Feeding Taiwan Loach

In this experiment, brewer’s yeast was used to replace part of the fish meal in the feed to formulate a high-fat and low-protein feed for Taiwan loach. The effects of brewer’s yeast on the growth performance, muscle quality, hepatopancreas and pancreas carnitine content, intestinal flora, immunity, and antioxidant ability of Taiwan loach were preliminarily investigated. In experiment 1, 600 Taiwan loaches were randomly divided into 4 groups, and 1% (group A), 4% (group B), 8% (group C), and 12% (group D) of brewer’s yeast was substituted for an equal amount of fishmeal in the basal diets. The loaches in each group were fed for 60 d. In experiment 2, 60 Taiwan loaches were selected and randomly divided into a control group (Group E) and a test group (Group F). The loaches were fed the basal diet and the high-fat and low-protein diet supplemented with 8% fishmeal, replaced by brewer’s yeast. The experiment lasted for 60 days, and the growth performance, hepatopancreas carnitine content, muscle quality, intestinal flora, plasma antioxidant, and immune capacity indices of loaches in each group were determined. The results were as follows: the weight gain rate and specific growth rate were significantly higher in group C than those in groups A, B, and D (P &lt; 0.05); the intestinal length ratio, muscle hardness, hepatopancreatic carnitine content, plasma superoxide dismutase activity, catalase activity, total antioxidant capacity, and lysozyme activity were significantly greater in group F than those in group E (P &lt; 0.05); the Lactobacillus and Bifidobacterium counts in Group F were considerably higher than those in Group E (P &lt; 0.05), and the Salmonella counts in Group F was significantly lower than that in Group E (P &lt; 0.05); the survival rate, weight gain rate, bait coefficient, muscle crude protein, crude fat, viscosity, elasticity, cohesion, chewability, and restorative capacity were not significantly different between Group F and Group E (P &gt; 0.05). The above results showed that adding 8% brewer’s yeast to feed Taiwan loach instead of fish meal could enhance its hepatopancreatic carnitine synthesis, optimize the intestinal flora, improve the body’s immune and antioxidant ability, and have a certain promotional effect on the intestinal development and muscle quality improvement.

Vitamin B5 supplementation enhances intestinal development and alters microbes in weaned piglets

This study explored the effects of different vitamin B5 (VB5) levels on intestinal growth and function of weaned piglets. Twenty-one piglets (7.20 ± 1.11 kg) were included in a 28-day feeding trial with three treatments, including 0 mg/kg (L-VB5), 10 mg/kg (Control) and 50 mg/kg (H-VB5) of VB5 supplement. The results showed that: Large intestine weight/body weight was the highest in H-VB5 group, Control and H-VB5 groups had significantly higher villus height and villus height/crypt depth than the L-VB5 in the ileum (p < .05). Goblet cells (ileal crypt) and endocrine cells (ileal villus) significantly increased in Control and H-VB5 (p < .05). The H-VB5 group exhibited significantly higher levels of ki67 and crypt depth in the cecum and colon, colonic goblet cells and endocrine cells were both rising considerably (p < .05). Isobutyric acid and isovaleric acid were significantly reduced in the H-VB5 group (p < .05), and there was a decreasing trend in butyric acid (p = .073). At the genus level, the relative abundance of harmful bacteria such as Clostridium_Sensu_Structo_1 Strecto_1, Terrisporbacter and Streptococcus decreased significantly and the relative abundance of beneficial bacteria Turicibacter increased significantly in H-VB5 group (p < .05). Overall, the addition of 50 mg/kg VB5 primarily enhanced the morphological structure, cell proliferation and differentiation of the ileum, cecum and colon. It also had a significant impact on the gut microbiota and short-chain fatty acids.

The secreted protein Amuc_1409 from Akkermansia muciniphila improves gut health through intestinal stem cell regulation

Akkermansia muciniphila has received great attention because of its beneficial roles in gut health by regulating gut immunity, promoting intestinal epithelial development, and improving barrier integrity. However, A. muciniphila-derived functional molecules regulating gut health are not well understood. Microbiome-secreted proteins act as key arbitrators of host-microbiome crosstalk through interactions with host cells in the gut and are important for understanding host-microbiome relationships. Herein, we report the biological function of Amuc_1409, a previously uncharacterised A. muciniphila-secreted protein. Amuc_1409 increased intestinal stem cell (ISC) proliferation and regeneration in ex vivo intestinal organoids and in vivo models of radiation- or chemotherapeutic drug-induced intestinal injury and natural aging with male mice. Mechanistically, Amuc_1409 promoted E-cadherin/β-catenin complex dissociation via interaction with E-cadherin, resulting in the activation of Wnt/β-catenin signaling. Our results demonstrate that Amuc_1409 plays a crucial role in intestinal homeostasis by regulating ISC activity in an E-cadherin-dependent manner and is a promising biomolecule for improving and maintaining gut health.

Generation of a Bovine Primary Enteroid-Derived Two-Dimensional Monolayer Culture System for Applications in Translational Biomedical Research.

Organoid cell culture systems can recapitulate the complexity observed in tissues, making them useful in studying host-pathogen interactions, evaluating drug efficacy and toxicity, and tissue bioengineering. However, applying these models for the described reasons may be limited because of the three-dimensional (3D) nature of these models. For example, using 3D enteroid culture systems to study digestive diseases is challenging due to the inaccessibility of the intestinal lumen and its secreted substances. Indeed, stimulation of 3D organoids with pathogens requires either luminal microinjection, mechanical disruption of the 3D structure, or generation of apical-out enteroids. Moreover, these organoids cannot be co-cultured with immune and stromal cells, limiting in-depth mechanistic analysis into pathophysiological dynamics. To circumvent this, we optimized a bovine primary cell two-dimensional (2D) enteroid-derived monolayer culture system, allowing co-culture with other relevant cell types. Ileal crypts isolated from healthy adult cattle were cultured to generate 3D organoids that were cryopreserved for future use. A 2D monolayer was created using revived 3D enteroids that were passaged and disrupted to yield single cells, which were seeded on basement membrane extract-coated transwell cell culture inserts, thereby exposing their apical surface. The intestinal monolayer polarity, cellular differentiation, and barrier function were characterized using immunofluorescence microscopy and measuring transepithelial electrical resistance. Stimulation of the apical surface of the monolayer revealed the expected functionality of the monolayer, as demonstrated by cytokine secretion from both apical and basal compartments. The described 2D enteroid-derived monolayer model holds great promise in investigating host-pathogen interactions and intestinal physiology, drug development, and regenerative medicine.

Effect of Ginger Root Extract on Intestinal Oxidative Status and Mucosal Morphometrics in Broiler Chickens.

This study was designed to assess the effect of ginger root extract (GRE) supplementation on the oxidative status and intestinal mucosal development in broiler chickens for 6 weeks. Day-old chicks (Ross 708 strain, n = 432) were distributed into six treatments with six replicate of twelve birds each: Negative CON (basal), MX (basal diet + bacitracin methylene disalicylate (BMD) 0.055 g/kg diet), GRE-1 (basal diet + 0.375% GRE), GRE-2 (basal diet + 0.75% GRE), GRE-3 (basal diet + 1.5% GRE), GRE-4 (basal diet + 3% GRE). Growth indices, goblets cell count, mucin (MUC2) in ileum tissue, antioxidant (SOD, CAT, and GPX) in ileum and liver, biological antioxidant potential (BAP), and reactive oxygen metabolite level in blood and intestinal villi measurement were determined. Body weight (BW) was highest (p < 0.05) in all groups except GRE-4, body weight gain (BWG) was best in GRE-1, while FCR was least in all groups except GRE-4. Optimum MUC2 gene expression, SOD, CAT, blood antioxidants, and intestinal morphometric values were observed in GRE-3. The inclusion of ginger root extract up to 1.5% improved growth and reduced oxidative stress while enhancing mucosal development in broiler chicks.

Microencapsulated Lactobacillus plantarum promotes intestinal development through gut colonization of layer chicks

The effects of Lactobacillus plantarum in microencapsulation (LPM) on intestinal development in layer chicks were investigated in this study, as well as the colonization of L. plantarum in the gut. A total of 480 healthy Hy-Line Brown layer chicks at 0 d old were randomly divided into 4 groups (8 replicates each treatment), and the diets of these birds were supplemented with nothing, L. plantarum, LPM and wall material of LPM (WM), respectively. LPM improved growth performance and intestinal development of layer chicks, evidenced by significantly increased body weight, average daily gain, average daily feed intake, villus height, villus height/crypt depth, as well as weight and length of the duodenum, jejunum and ileum (P ˂ 0.05). These results could be attributed to the increased colonization of L. plantarum in the gut, which was verified by significant increases in lactic acid content, viable counts in chyme and mucosa (P ˂ 0.05), as well as a visible rise in number of strains labeled with fluorescein isothiocyanate. Meanwhile, the relative abundances of Lactobacillus and Bifidobacterium significantly increased in response to microencapsulated L. plantarum supplementation (P ˂ 0.05), accompanied by the significant up-regulation of colonization related genes (P ˂ 0.05), encoding solute carrier family, monocarboxylate transporter, activin A receptor, succinate receptor and secretogranin Ⅱ. To sum up, microencapsulated L. plantarum supplementation promoted intestinal development, which could be attributed to the enhancement of L. plantarum colonization in the intestine through the mutual assistance of Bifidobacterium and interactions with colonization related transmembrane proteins.

Interferon regulatory factor 6 (IRF6) determines intestinal epithelial cell development and immunity

Intestinal epithelial cell (IEC) responses to interferon (IFN) favor antiviral defense with minimal cytotoxicity, but IEC-specific factors that regulate these responses remain poorly understood. Interferon regulatory factors (IRFs) are a family of nine related transcription factors, and IRF6 is preferentially expressed by epithelial cells, but its roles in IEC immunity are unknown. In this study, CRISPR screens found that Irf6 deficiency enhanced IFN-stimulated antiviral responses in transformed mouse IECs but not macrophages. Furthermore, KO ofIrf6in IEC organoids resulted in profound changes to homeostasis and immunity gene expression. Irf6 KO organoids grew more slowly, and single-cell RNA sequencing indicated reduced expression of genes in epithelial differentiation and immunity pathways. IFN-stimulated gene expression was also significantly different inIrf6KO organoids, with increased expression of stress and apoptosis-associated genes. Functionally, the transcriptional changes in Irf6 KO organoids were associated with increased cytotoxicity upon IFN treatment or inflammasome activation. These data indicate a previously unappreciated role for IRF6 in IEC biology, including regulation of epithelial development and moderation of innate immune responses to minimize cytotoxicity and maintain barrier function.

Interactions between intestinal microbiota and metabolites in zebrafish larvae exposed to polystyrene nanoplastics: Implications for intestinal health and glycolipid metabolism

Previous studies have shown the harmful effects of nanoscale particles on the intestinal tracts of organisms. However, the specific mechanisms remain unclear. Our present study focused on examining the uptake and distribution of polystyrene nanoplastics (PS-NPs) in zebrafish larvae, as well as its toxic effects on the intestine. It was found that PS-NPs, marked with red fluorescence, primarily accumulated in the intestine section. Subsequently, zebrafish larvae were exposed to normal PS-NPs (0.2–25 mg/L) over a critical 10-day period for intestinal development. Histopathological analysis demonstrated that PS-NPs caused structural changes in the intestine, resulting in inflammation and oxidative stress. Additionally, PS-NPs disrupted the composition of the intestinal microbiota, leading to alterations in the abundance of bacterial genera such as Pseudomonas and Aeromonas, which are associated with intestinal inflammation. Metabolomics analysis showed alterations in metabolites that are primarily involved in glycolipid metabolism. Furthermore, MetOrigin analysis showed a significant correlation between bacterial flora (Pedobacter and Bacillus) and metabolites (D-Glycerate 2-phosphate and D-Glyceraldehyde 3-phosphate), which are related to the glycolysis/gluconeogenesis pathways. These findings were further validated through alterations in multiple biomarkers at various levels. Collectively, our data suggest that PS-NPs may impair the intestinal health, disrupt the intestinal microbiota, and subsequently cause metabolic disorders.

Dietary apidaecin Api-PR19 addition improves growth performance, intestinal health and modulating gut microbiota in broilers.

This study investigated the effects of Apidaecin Api-PR19 as feed additive on growth performance, intestinal health, and small intestinal microbiota of broilers. A total of 360 1-d-old Arbor Acres broilers were randomly assigned to 3 groups with 6 replicates including control group with basal diet (CON), antibiotic growth promotor group with basal plus 10 mg/kg colistin sulfate and 50mg/kg roxarsone (AGP), and antibacterial peptide group with basal diet plus 330 mg/kg Apidaecin Api-PR19 (ABP). The trial lasted 35 d. Results showed that dietary Api-PR19 addition increased (p＜0.05) the average daily feed intake (ADFI), average daily gain (ADG) and decreased (p＜0.05) feed conversion ratio (FCR) during 1 to 21 d compared with the CON group. The digestibility of dry matter and crude protein were higher in AGP and ABP groups (p＜0.05) where greater trypsin activity was detected in duodenum (p＜0.05). The ratio of villus height to crypt depth (V/R) in duodenum and jejunum was increased at 35 d when broilers were given diets with ABP or AGP (p＜0.05). Besides, ABP treatments up-regulated (p＜0.05) the mRNA expression of EAAT3, GLUT2, ZO-1 and Claudin-1 in duodenum of broilers at 35 d of age. The results of immunohistochemistry showed that ABP treatment significantly increased (p＜0.05) duodenal sIgA content. In addition, 16S rRNA gene sequencing revealed that there were differences in the intestinal microbiota diversity and composition among three groups. Notably, the linear discriminant analysis effect size (LEfSe) showed that p_Firmicutes, g_Enterococcus, g_Carnobacterium, g_Kitasatospora and g_Acidaminococcus were dominant in ABP group. Redundancy analysis showed that these changes in gut microbiota in ABP group had correlation with growth performance, intestinal morphology, and content of sIgA. In general, these results indicated that dietary 330 mg/kg Apidaecin Api-PR19 supplementation promoted growth performance of broilers by improving intestinal development, nutrients absorption, immune function and modulating intestinal microbiota.