Pig Intestine Research Articles

Selection and characterization of bacteriophages specific to Salmonella Choleraesuis in swine.

Salmonella Choleraesuis is the most common serotype that causes salmonellosis in swine. Recently, the use of bacteriophages as a potential biocontrol strategy has increased. Therefore, this study aimed to isolate and characterize bacteriophages specific to S. Choleraesuis associated with swine infection and to evaluate the efficacy of individual phages and a phage cocktail against S. Choleraesuis strains in simulated intestinal fluid (SIF). Three strains of S. Choleraesuis isolated from pig intestines served as host strains for phage isolation. The other 10 Salmonella serovars were also used for the phage host range test. The antibiotic susceptibility of the bacterial strains was investigated. Water samples from natural sources and drain liquid from slaughterhouses were collected for phage isolation. The isolated phages were characterized by determining the efficiency of plating against all Salmonella strains and the stability at a temperature range (4°C-65°C) and at low pH (2.5-4.0) in simulated gastric fluids (SGFs). Furthermore, morphology and genomic restriction analyses were performed for phage classification phages. Finally, S. Choleraesuis reduction in the SIF by the selected individual phages and a phage cocktail was investigated. The antibiotic susceptibility results revealed that most Salmonella strains were sensitive to all tested drugs. Salmonella Choleraesuis KPS615 was multidrug-resistant, showing resistance to three antibiotics. Nine phages were isolated. Most of them could infect four Salmonella strains. Phages vB_SCh-RP5i3B and vB_SCh-RP61i4 showed high efficiency in infecting S. Choleraesuis and Salmonella Rissen. The phages were stable for 1 h at 4°C-45°C. However, their viability decreased when the temperature increased to 65°C. In addition, most phages remained viable at a low pH (pH 2.5-4.0) for 2 h in SGF. The efficiency of phage treatment against S. Choleraesuis in SIF showed that individual phages and a phage cocktail with three phages effectively reduced S. Choleraesuis in SIF. However, the phage cocktails were more effective than the individual phages. These results suggest that the newly isolated phages could be promising biocontrol agents against S. Choleraesuis infection in pigs and could be orally administered. However, further in vivo studies should be conducted.

Intraepithelial lymphocytes in the pig intestine: T cell and innate lymphoid cell contributions to intestinal barrier immunity.

Intraepithelial lymphocytes (IELs) include T cells and innate lymphoid cells that are important mediators of intestinal immunity and barrier defense, yet most knowledge of IELs is derived from the study of humans and rodent models. Pigs are an important global food source and promising biomedical model, yet relatively little is known about IELs in the porcine intestine, especially during formative ages of intestinal development. Due to the biological significance of IELs, global importance of pig health, and potential of early life events to influence IELs, we collate current knowledge of porcine IEL functional and phenotypic maturation in the context of the developing intestinal tract and outline areas where further research is needed. Based on available findings, we formulate probable implications of IELs on intestinal and overall health outcomes and highlight key findings in relation to human IELs to emphasize potential applicability of pigs as a biomedical model for intestinal IEL research. Review of current literature suggests the study of porcine intestinal IELs as an exciting research frontier with dual application for betterment of animal and human health.

BACTERIAL INTESTINAL INFECTIONS OF SWINE

Bacterial intestinal infections are one of the main diseases in pigs of different ages. Bacterial diseases of the pig digestive tract lead to significant economic losses due to high mortality, reduced growth, treatment and prevention costs.
 The main bacterial intestinal infections of pigs are anaerobic enterotoxemia (clostridiosis), colibacillosis, intestinal salmonellosis, dysentery, proliferative enteropathy (ileitis).
 Anaerobic enterotoxemia of pigs is an acute toxic-infectious disease mainly of newborn piglets, caused by pathogenic bacteria of the genus Clostridium and characterized by hemorrhagic-necrotic inflammation of the intestinal mucosa, diarrhea and toxicosis.
 Swine colibacillosis is an intestinal infection caused by the enterotoxigenic Escherichia coli, which is able to produce enterotoxins that locally affect the intestines of pigs, causing diarrheal syndrome.
 Intestinal salmonellosis is a factorial infection. The causative agents are enteropathogenic salmonella (mainly Salmonella enterica serotype typhimurium), which cause inflammation and necrosis of the small and large intestine, leading to diarrhea, which may be accompanied by generalized sepsis.
 Dysentery is a severe enteroinfection of pigs caused by the anaerobic bacterium Brachyspira hyodysenteriae, characterized by fever, debilitating mucohemorrhagic diarrhea and dehydration, leading to high mortality among animals.
 Proliferative enteropathy is a sporadic disease of pigs caused by Lawsonia intracellularis. The acute form of ileitis, known as proliferative hemorrhagic enteritis, is characterized by intestinal hemorrhage and sudden death, and usually occurs in pigs older than 4 months.
 Diarrhea in piglets can also be caused by enterococci (Enterococcus spp.) and chlamydia (Chlamydia suis). Enterococcal bacteria cause diarrhea in newborn piglets, and intestinal chlamydia infections are mostly common in rearing piglets, and it is believed that most intestinal infections caused by chlamydia are subclinical.
 The literature review regarding the etiology, pathogenesis and clinical diagnosis of major bacterial intestinal infections in pigs is presented in the article.

Magnetically Controlled Multifunctional Capsule Robot for Dual-Drug Delivery

This article presents a multifunctional capsule robot (MCR) with active locomotion, dual-drug load, and selective drug release. The MCR has a high ratio of the volume of loaded drug to the total volume of the capsule (RDC) and can provide sufficient dosages for treatment. The MCR might be used to address the limitations in robot-assisted drug delivery, greatly improving the efficiency of intestinal disease treatment. The MCR is composed of a locomotion unit and a drug delivery unit, and these units are controlled by two orthogonal rotating magnetic fields. This proposed MCR enables active locomotion to target positions and selects to release two different drugs according to clinic requirements. <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">In vitro</i> and <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">ex vivo</i> experiments were conducted to evaluate the performance of the MCR. Experimental results show that the maximum drug loading capacity of the MCR is 1.5 g, and the maximum advance and retreat velocities are 11.8 mm/s and 10.4 mm/s, respectively, when the MCR is fully loaded. The MCR can release two different drugs in various targets and no noticeable damages to the pig intestine were observed to the naked eyes. The MCR will have potential further application for intestinal disease treatment.

Metagenomics analysis reveals potential pathways and drivers of piglet gut phage-mediated transfer of ARGs

The growing prevalence of antibiotic-resistant pathogens has led to a better understanding of the underlying processes that lead to this expansion. Intensive pig farms are considered one of the hotspots for antibiotic resistance gene (ARG) transmission. Phages, as important mobile carriers of ARGs, are widespread in the animal intestine. However, our understanding of phage-associated ARGs in the pig intestine and their underlying drivers is limited. Here, metagenomic sequencing and analysis of viral DNA and total DNA of different intestinal (ileum, cecum and feces) contents in healthy piglets and piglets with diarrhea were separately conducted. We found that phages in piglet ceca are the main repository for ARGs and mobile genetic element (MGE) genes. Phage-associated MGEs are important factors affecting the maintenance and transfer of ARGs. Interestingly, the colocalization of ARGs and MGE genes in piglet gut phages does not appear to be randomly selected but rather related to a specific phage host (Streptococcus). In addition, in the feces of piglets with diarrhea, the abundance of phages carrying ARGs and MGE genes was significantly increased, as was the diversity of polyvalent phages (phages with broad host ranges), which would facilitate the transfection and wider distribution of ARGs in the bacterial community. Moreover, the predicted host spectrum of polyvalent phages in diarrheal feces tended to be potential enteropathogenic genera, which greatly increased the risk of enteropathogens acquiring ARGs. Notably, we also found ARG-homologous genes in the sequences of piglet intestinal mimiviruses, suggesting that the piglet intestinal mimiviruses are a potential repository of ARGs. In conclusion, this study greatly expands our knowledge of the piglet gut microbiome, revealing the underlying mechanisms of maintenance and dissemination of piglet gut ARGs and providing a reference for the prevention and control of ARG pollution in animal husbandry.

Development of Shelf Stable Chitterlings Pickle Incorporated with Natural Acidulant

Background: Intestine of pigs (chitterling) is one of the important edible offal with substantial yield and accounts for 1.8% of the slaughter weight. At present its utilization as direct consumption is limited only to some economically weaker section of population. Therefore the present study was undertaken to develop a shelf stable chitterling pickle recipe incorporated with fermented bamboo shoot (FBS) and lemon juice as natural acidulant. Methods: Freshly collected intestines of pig were washed, pressure cooked, cut into 3 to 4 cm3 and deep fried. Spices and condiments were fried separately then mixed with the fried chitterlings along with FBS and lemon juice to prepare the shelf stable chitterling pickle. The pickles were studied for various parameters such as physico-chemical, microbial and sensory attributes. Result: Our study showed significantly (p less than 0.05) higher pH, product yield and moisture content in the chitterling pickle added with FBS and lemon juice. Protein and fat content did not differ significantly. No adverse effects were noted on hedonic score for appearance, flavor, juiciness, texture, saltiness, sourness and overall acceptability up to 12 days of storage at ambient temperature. The overall acceptability of chitterling pickle with FBS and lemon juice was significantly (p less than 0.05) higher as compared to control. Therefore, it could be concluded that highly acceptable shelf stable pickle can be prepared from intestine of pig with substantial value addition to the by-products.

Artificial blood-feeding of phlebotomines (Diptera: Psychodidae: Phlebotominae): is it time to repurpose biological membranes in light of ethical concerns?

BackgroundThe aims of the present study were to evaluate and compare the efficacy of blood-feeding in phlebotomines through industrially processed membranes from the small intestine of pigs (used for the production of commercial sausages) and the skin of euthanized chicks.MethodsLaboratory-bred Lutzomyia longipalpis and different field-caught phlebotomine species were subjected to the artificial feeding systems under similar conditions. Paired tests were performed using the control (skin from euthanized chicks) and test membranes (pig small intestine). The feeding rates were compared by paired t-test, and Pearson correlation was used to examine the relationship between the thickness of the membranes and feeding rate.ResultsThe feeding rate was greater with the test membrane than with the control membrane for L. longipalpis (t-test, t = −3.3860, P = 0.0054) but not for the most frequent field-caught species, Nyssomyia antunesi (t-test, t = 0.7746, P = 0.4535). The average thicknesses of the control and test membranes were 184 ± 83 µm and 34 ± 12 µm, respectively (Mann–Whitney U-test, U = 0.00, Z = 2.8823, P = 0.0039); however, there was no correlation between feeding rate and membrane thickness. A moderate positive correlation was observed between the number of phlebotomines that fed and the total number of phlebotomines in the cage for each type of membrane and for each species.ConclusionsThe test membrane is a viable alternative for the artificial blood-feeding of phlebotomines, and is thus a potential substitute for the skin of animals that are euthanized for this purpose. Feeding rate was independent of membrane thickness.Graphical

Regional epithelial cell diversity in the small intestine of pigs.

Understanding regional distribution and specialization of small intestinal epithelial cells is crucial for developing methods to control appetite, stress, and nutrient uptake in swine. To establish a better understanding of specific epithelial cells found across different regions of the small intestine in pigs, we utilized single-cell RNA sequencing (scRNA-seq) to recover and analyze epithelial cells from duodenum, jejunum, and ileum. Cells identified included crypt cells, enterocytes, BEST4 enterocytes, goblet cells, and enteroendocrine (EE) cells. EE cells were divided into two subsets based on the level of expression of the EE lineage commitment gene, NEUROD1. NEUROD1hi EE cells had minimal expression of hormone-encoding genes and were dissimilar to EE cells in humans and mice, indicating a subset of EE cells unique to pigs. Recently discovered BEST4 enterocytes were detected in both crypts and villi throughout the small intestine via in situ staining, unlike in humans, where BEST4 enterocytes are found only in small intestinal villi. Proximal-to-distal gradients of expression were noted for hormone-encoding genes in EE cells and nutrient transport genes in enterocytes via scRNA-seq, demonstrating regional specialization. Regional gene expression in EE cells and enterocytes was validated via quantitative PCR (qPCR) analysis of RNA isolated from epithelial cells of different small intestinal locations. Though many genes had similar patterns of regional expression when assessed by qPCR of total epithelial cells, some regional expression was only detected via scRNA-seq, highlighting advantages of scRNA-seq to deconvolute cell type-specific regional gene expression when compared to analysis of bulk samples. Overall, results provide new information on regional localization and transcriptional profiles of epithelial cells in the pig small intestine.

NABT

NABT

Preclinical evaluation of a veno-venous bypass device for liver transplantation based on the principle of magnetic levitation drive

Objective: To explore the performance of a self-made venous-venous bypass (VVB) device for liver transplantation based on the principle of magnetic levitation drive. Methods: Experimental study was conducted from August 2020 to January 2022. Eight Bama minipigs underwent VVB of hepatic portal vein-femoral vein-internal jugular vein after occlusion of hepatic portal vein and inferior vena cava. The animals were divided into two groups according to the VVB devices used during VVB. A self-made VVB device was used in group A(n=5),and an imported VVB device was used in group B(n=3). The hemodynamic changes of the two groups of animals were compared at 6 time points including before vascular occlusion, during vascular occlusion, 30 minutes, 60 minutes, 90 minutes after the start of VVB, and 30 minutes after vascular opening. In addition,the changes of blood compatibility indexes,intestinal injury indexes,kidney injury indexes and internal environment indexes of the two groups of animals at each time point were compared. The independent samples t test was used for the quantitative data between the two groups with non-repeated measures,and the repeated measures analysis of variance was used for the quantitative data between the two groups with repeated measures. Results: During the VVB of the two devices,the venous drainage was sufficient,and the main manifestations were that the color of the intestine of the Bama miniature pig was ruddy, the peristalsis was normal, and the urine output was normal. There were no significant differences in hemodynamics,blood injure indexes,intestinal injury indexes,kidney injury indexes,neutropil gelatinase-associated lipocalin,and internal environment indexes(all P>0.05).The indexes at 30 minutes after vascular opening in the group A and the group B were as follows:mean arterial pressure were (71.0±7.7)mmHg(1 mmHg=0.133 kPa) and (74.0±8.7)mmHg,central venous pressure were (7.0±1.4)cmH2O(1 cmH2O=0.098 kPa) and (7.7±0.6)cmH2O,heart rate were (131±10) beats/minutes and (132±8)beats/minutes; red blood cell count were (6.43±0.89)×1012/L and (6.32±0.58)×1012/L,hemoglobin were (108.4±5.9)g/L and (110.0±3.5)g/L,free hemoglobin were (78.28±3.96)mg/L and (78.08±4.54)mg/L; intestinal fatty acid binding protein were (2.27±0.49)μg/L and (2.40±0.78)μg/L;creatinine were (68.30±9.77)μmol/L and (79.90±26.91)μmol/L,blood urea nitrogen were (3.94±1.39)mmol/L and (3.45±0.65)mmol/L;neutropil gelatinase-associated lipocalin were (4.02±0.53) μg/L and (3.86±0.23)μg/L;pH value were 7.27±0.04 and 7.23±0.03,lactic acid were (6.18±2.62)mmol/L and (4.30±0.50)mmol/L,concentrations of Na+ were (136.3±3.0)mmol/L and (137.6±1.6) mmol/L,concentrations of K+ were (3.89±0.42) mmol/L and (3.98±0.17)mmol/L,concentrations of Ca2+ were (1.40±0.03)mmol/L and(1.40±0.04)mmol/L;all indexes in the two group had no differences(all P>0.05). Conclusion: The self-made venous bypass device can be safely and effectively applied to VVB of Bama minipigs,and achieves the same performance as the imported venous bypass device.

Complete genomic sequence analysis and intestinal tissue localization of a porcine Kobuvirus variant in China.

Porcine kobuvirus (PKV) infection is very common in both healthy pigs and diarrhea pigs throughout the world. However, there is no proof that it causes diarrhea, and little is known about its role in diarrhea. There are only a few reports concerning porcine kobuvirus separation at present, which makes investigating its invasion and pathogenesis mechanisms difficult. This study sequenced the entire genome of a porcine kobuvirus strain termed "Wuhan2020" after it was isolated from intestinal tissue samples of healthy piglets. The analysis results revealed that it shared the most resemblance with the WUH1 strain (89.5%) and belonged to the same evolutionary branch as the Hungarian strain S-1-SUN. The PKV was located using the in situ hybridization (ISH) approach, which revealed that it was colonized in intestinal villus epithelial cells and lymphocytes in the Peyer's patch. In general, we analyzed the genetic evolution of PKV, discovered PKV susceptible cells and determined PKV localization in the intestine of infected pigs, providing a reference for future research.

A microexplosive shockwave-based drug delivery microsystem for treating hard-to-reach areas in the human body

Implantable drug-delivery microsystems have the capacity to locally meet therapeutic requirements by maximizing local drug efficacy and minimizing potential side effects. The internal organs of the human body including the esophagus, gastrointestinal tract, and respiratory tract, with anfractuos contours, all manifest with endoluminal lesions often located in a curved or zigzag area. The ability of localized drug delivery for these organs using existing therapeutic modalities is limited. Spraying a drug onto these areas and using the adhesion and water absorption properties of the drug powder to attach to lesion areas can provide effective treatment. This study aimed to report the development and application of microsystems based on microshockwave delivery of drugs. The devices comprised a warhead-like shell with a powder placed at the head of the device and a flexible rod that could be inserted at the tail. These devices had the capacity to deposit drugs on mucous membranes in curved or zigzag areas of organs in the body. The explosive impact characteristics of the device during drug delivery were analyzed by numerical simulation. In the experiment of drug delivery in pig intestines, we described the biosafety and drug delivery capacity of the system. We anticipate that such microsystems could be applied to a range of endoluminal diseases in curved or zigzag regions of the human body while maximizing the on-target effects of drugs.

272 Effects of a Probiotic Bacillus Strain on Ileal Digestibility of Crude Protein, Starch, Energy and fat and Total Tract Digestibility of Energy and Dietary Fiber in Diets fed to Weanling Pigs

Abstract An experiment was conducted to test the hypothesis that probiotic Bacillus toyonensis M15750 improve the apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of energy and nutrients when included in diets fed by weanling pigs. A control diet was formulated based on corn, soybean-meal, and distillers dried grains with solubles and a second diet was formulated by supplementing the probiotic Bacillus toyonensis M15750 (1 × 109 cfu/kg feed) to the control diet. Titanium dioxide (0.50 %) was added to the diets as an indigestible marker. Sixteen weanling barrows (10.7 ± 0.7 kg) that had a T-cannula in the distal ileum were allotted to the two diets with 8 replicate pigs per diet in a randomized complete block design with body weight being the blocking factor. Pigs were adapted to the diets and to the metabolism crates for 7 days, urine and fecal materials were collected during the following 4 days according to standard procedures using the marker-to-marker approach, whereas ileal digesta were collected for 8 h during the following 2 days. The AID and ATTD of energy and nutrients were calculated for each diet. The AID of dry matter and gross energy was greater (P &amp;lt; 0.05) and the AID of starch tended to be greater (P = 0.05) in the diet supplemented with the probiotic Bacillus toyonensis M15750 compared with the control diet (Table 1). However, no differences were observed for the ATTD of energy or nutrients between the 2 diets, which indicates that the increased digestibility in the small intestine of pigs fed the diet supplemented with Bacillus toyonensis M15750 was offset by a reduction in hindgut fermentation. These results indicate that inclusion of probiotic Bacillus toyonensis M15750 in diets for weanling pigs have the potential to improve the AID of dry matter, gross energy and starch.

Extraction, purification, and in vitro biological activities of intestinal alkaline phosphatase from pig intestine mucous waste

Extraction, purification, and in vitro biological activities of intestinal alkaline phosphatase from pig intestine mucous waste

Lactiplantibacillus plantarum Regulated Intestinal Microbial Community and Cytokines to Inhibit Salmonella typhimurium Infection.

Lactic acid bacteria (LAB) are recognized as food-grade safe microorganisms and have many beneficial effects. LAB could maintain the host intestinal homeostasis and regulate intestinal microbial community to exert antibacterial effects. In this study, Lactiplantibacillus plantarum (L. plantarum, Lp01) strain isolated from pig intestine was orally administered to C57BL/6 mice, and mice were then infected with Salmonella typhimurium (ATCC14028). The protective effects of L. plantarum were evaluated by monitoring body weight loss, survival rates, bacterial loads in tissue, colon histopathology analysis, and cytokine secretion. 16S rRNA gene sequencing was also utilized to detect the dynamics of the blind gut microbial community in mice. We found that L. plantarum could significantly reduce the body weight loss and improve the survival rates. The survival rate in the L. P-Sty group was up to 67.5%, which was much higher than that in the STY group (25%). Counting of bacterial loads displayed that the colony-forming unit (CFU) of S. typhimurium in the spleen (p < 0.05) and the liver (p < 0.05) from L. P-Sty group both decreased, compared with STY group. Intestinal histopathology showed that it alleviated the intestinal injury caused by Salmonella, inhibited the secretion of pro-inflammatory cytokines, and promoted anti-inflammatory cytokines (p < 0. 01). In addition, L. plantarum also significantly ameliorated the intestinal gut microbiome disturbance caused by Salmonella. It displayed an obvious increase of beneficial bacteria including Lactobacillus and Bacteroidetes and reduction of pathogenic bacteria like Proteobacteria. In conclusion, L. plantarum could regulate microbial community to inhibit Salmonella typhimurium infection.

Lupinus albus γ-Conglutin: New Findings about Its Action at the Intestinal Barrier and a Critical Analysis of the State of the Art on Its Postprandial Glycaemic Regulating Activity

γ-Conglutin (γ-C) is the glycoprotein from the edible seed L. albus, studied for long time for its postprandial glycaemic regulating action. It still lacks clear information on what could happen at the meeting point between the protein and the organism: the intestinal barrier. We compared an in vitro system involving Caco-2 and IPEC-J2 cells with an ex vivo system using pig ileum and jejunum segments to study γ-C transport from the apical to the basolateral compartment, and its effects on the D-glucose uptake and glucose transporters protein expression. Finally, we studied its potential in modulating glucose metabolism by assessing the possible inhibition of α-amylase and α-glucosidase. RP-HPLC analyses showed that γ-C may be transported to the basolateral side in the in vitro system but not in the pig intestines. γ-C was also able to promote a decrease in glucose uptake in both cells and jejunum independently from the expression of the SGLT1 and GLUT2 transporters.

Three-Dimensional Structure of the Antimicrobial Peptide Cecropin P1 in Dodecylphosphocholine Micelles and the Role of the C-Terminal Residues.

Cecropin P1 (CP1) isolated from a large roundworm Ascaris suum, which is found in pig intestines, has been extensively studied as a model antimicrobial peptide (AMP). However, despite being a model AMP, its antibacterial mechanism is not well understood, particularly the function of its C-terminus. By using an Escherichia coli overexpression system with calmodulin as a fusion partner, we succeeded in the mass expression of recombinant peptides, avoiding toxicity to the host and degradation of CP1. The structure of the recombinant 15N- and 13C-labeled CP1 and its C-terminus truncated analogue in dodecylphosphocholine (DPC) micelles was determined by NMR. In this membrane-mimetic environment, CP1 formed an α-helix for almost its entire length, except for a short region at the C-terminus, and there was no evidence of a hinge, which is considered important for the expression of activity in other cecropins. Several NMR analyses showed that the entire length of CP1 was protected from water by micelles. Since the loss of the C-terminus of the analogue had little effect on the NMR structure or its interaction with the micelle, we investigated another role of the C-terminus of CP1 in its antimicrobial activity. The results showed that the C-terminal region affected the DNA-binding capacity of CP1, and this mechanism of action was also newly suggested that it contributed to the antimicrobial activity of CP1.

Ammonia-induced excess ROS causes impairment and apoptosis in porcine IPEC-J2 intestinal epithelial cells

Ammonia is one of the most important toxic metabolites in the intestine of animals. It can cause intestinal damage and associated intestinal diseases through different endogenous or exogenous stimuli. However, the definition of harmful ammonia concentration and the molecular mechanism of ammonia - induced intestinal epithelial injury remain unclear. In this study, we found that the viability of porcine IPEC-J2 intestinal epithelial cells significantly decreased with the increase of NH4Cl dose (20–80 mM). Ammonia (40 mM NH4Cl) increased the expression level of ammonia transporter RHCG and disrupted the intestinal barrier function of IPEC-J2 cells by reducing the expression levels of the tight junction molecules ZO-1 and Claudin-1. Ammonia caused elevated levels of ROS and apoptosis in IPEC-J2 cells. This was manifested by decreased activity of antioxidant enzymes SOD and GPx, decreased mitochondrial membrane potential, and increased cytoplasmic Ca2+ concentration. In addition, the expression levels of apoptosis-related molecules Caspase-9, Caspase-3, Fas, Caspase-8, p53 and Bax were increased, the expression level of anti-apoptotic molecule Bcl-2 was decreased. Moreover, the antioxidant NAC (N-acetyl-L-cysteamine) effectively alleviated ammonia-induced cytotoxicity, reduced ROS level, Ca2+ concentration, and the apoptosis of IPEC-J2 cells. The results suggest that ammonia-induced excess ROS triggered apoptosis through mitochondrial pathway, death receptor pathway and DNA damage. This study can provide reference and theoretical basis for the definition of harmful ammonia concentration in pig intestine and the effect and mechanism of ammonia on pig intestinal health.

Importance of gut microbiota for bile acid composition and concentration in pigs

Several studies on the role of HCA species in regulating glucose homeostasis have indicated their therapeutic value in human obesity and diabetes. There is a clear difference in the percentage of hyocholic acid (HCA) and its derivatives (also known as HCA species) in the total bile acid (BA) pool in the plasma between humans, rats and pigs. However, the role of gut microbiota in BA profiles of pigs remains unclear. We generated five germ-free pig models and six gnotobiotic pig models by fecal microbiota transplantation (FMT). A total of 46 BAs were detected in the jejunum, cecum, colon, and rectum chyme, 37 and 33 BAs were detected in bile, 33 BAs were detected in ileal chyme and liver, and 31 BAs were detected in serum. FMT increased the percentages of HCA species in total bile acids in the serum (79%), liver (78%), and bile (71%), but decreased the proportions of HCA species in the total BAs of the ileum (61%), cecum (47%), colon (51%), and rectum (57%) of pigs, as compared to GF piglets. FMT significantly induced the production of conjugated bile acids in the small intestine and increased the concentrations of free BAs in the large intestine of pigs (P &amp;lt; 0.01). FMT piglets had over 68-fold and 104-fold increases in conjugated BAs in the ileum compared to the germ-free piglets. FMT piglets had an expression pattern distinct from that of germ-free piglets for genes involved in bile acid receptors, synthesis, signaling, and transport. The gene expression levels of the rate-limiting enzyme CYP7A1 and the enzymes CH25H and BAAT involved in BA synthesis were significantly decreased in the liver of FMT piglets, and there was a significant reduction in the gene expression of FXR and TGR5 through the FGFR4/β-Klotho pathway that promotes the BA pool in the liver of piglets after FMT.

Amino acid utilization allows intestinal dominance of Lactobacillus amylovorus.

The mammalian intestine harbors heterogeneous distribution of microbes among which specific taxa (e.g. Lactobacillus) dominate across mammals. Deterministic factors such as nutrient availability and utilization may affect microbial distributions. Due to physiological complexity, mechanisms linking nutrient utilization and the dominance of key taxa remain unclear. Lactobacillus amylovorus is a predominant species in the small intestine of pigs. Employing a pig model, we found that the small intestine was dominated by Lactobacillus and particularly L. amylovorus, and enriched with peptide-bound amino acids (PBAAs), all of which were further boosted after a peptide-rich diet. To investigate the bacterial growth dominance mechanism, a representative strain L. amylovorus S1 was isolated from the small intestine and anaerobically cultured in media with free amino acids or peptides as sole nitrogen sources. L. amylovorus S1 grew preferentially with peptide-rich rather than amino acid-rich substrates, as reflected by enhanced growth and PBAA utilization, and peptide transporter upregulations. Utilization of free amino acids (e.g. methionine, valine, lysine) and expressions of transporters and metabolic enzymes were enhanced simultaneously in peptide-rich substrate. Additionally, lactate was elevated in peptide-rich substrates while acetate in amino acid-rich substrates, indicating distinct metabolic patterns depending on substrate forms. These results suggest that an increased capability of utilizing PBAAs contributes to the dominance of L. amylovorus, indicating amino acid utilization as a deterministic factor affecting intestinal microbial distribution. These findings may provide new insights into the microbe-gut nutrition interplay and guidelines for dietary manipulations toward gut health especially small intestine health.