Pig Duodenum Research Articles

Humoral immune responses in piglets experimentally infected with a field strain of porcine epidemic diarrhea virus

Porcine epidemic diarrhea virus (PEDV) causes severe clinical diarrhea in neonatal piglets, with reported mortality rates between 70–100%. The humoral immunity, especially the local intestinal IgA responses, plays an important role in the immune protection against PEDV infection. In this study, we evaluated the isotype antibody responses against the PEDV nucleocapsid (N) protein and the spike (S) protein subunits 1 (S1) and 2 (S2) in the serum and intestine of piglets. We also determined its serum neutralizing activity against the PEDV field strain HBMC2012 in 21-day-old piglets. Enzyme-linked immunosorbent assays (ELISA) revealed that the production of IgM against the N protein and S1 subunit was higher compared to the S2 subunit. The anti-S2 IgA antibodies were higher than the anti-N protein and anti-S1 IgA at 3 days post-infection (dpi). The specific IgA responses to the S2 subunit were higher than the responses observed in S1. The specific IgG responses against S1 and S2 subunits exceeded those of N protein. The serum neutralizing activities against PEDV were relatively low with a tendency to decline over time. No isotype-specific antibodies were found in the intestinal contents from infected pigs, except the one with weak IgA responses against N protein at 28 dpi. Immunohistochemical staining showed that a few IgM, IgA, and IgG antibody-secreting cells were mainly located in the mucosa of the duodenum and ileum of PEDV-infected pigs at 3 dpi. This study suggests poor systemic and intestinal isotype-specific antibody responses, especially those of IgA, and weak serum neutralizing activities against the field PEDV strain in piglets.

Effect of Corn Particle Size on the Particle Size of Intestinal Digesta or Feces and Nutrient Digestibility of Corn-Soybean Meal Diets for Growing Pigs.

Simple SummaryDietary particle size can affect the digestible or metabolizable energy values of diet for pig. So, the mean particle size (MPS) of corn-soybean meal diet was recommended to less than 600 µm to improve growth performance and efficiency of dietary nutrients utilization by growing pigs. However, little information is available on whether further reduction of MPS influences the digestion process and increases digestibility of nutrients. This study evaluated the effects of reducing particle size of corn from 682 to 365 µm on the particle size of intestinal digesta or feces and nutrient digestibility of corn-soybean meal diets for growing pigs. The results showed that the MPS of duodenal digesta, ileal digesta and feces was reduced with the reduction of corn particle size. But, the MPS of corn did not affect the activities of amylase, trypsin and chymotrypsin in the duodenal fluid and the apparent total tract digestibility (ATTD) of dry matter, gross energy, crude protein, ether extract, neutral (NDF) and acid detergent fiber (ADF). These results suggest that the MPS less than 511 µm for corn-soybean meal diet does not improve the utilization of nutrients, thus it’s not necessary to further reduce particle size of corn with MPS of 682 µm.This study was conducted to evaluate the effect of corn particle size on the particle size of intestinal digesta or feces and nutrient digestibility of corn–soybean meal diets. Twenty-four growing barrows (initial BW: 21.9 ± 1.62 kg) were randomly divided into 4 groups of 6 pigs. A T-cannula was surgically placed in the anterior duodenum (about 50 cm from pylorus) of pigs in Groups 1 and 2 or in the distal ileum of pigs in Groups 3 and 4. Corn used to formulate diets had mean particle size (MPS) of 365 µm (Corn 1) or 682 µm (Corn 2), resulting in diets with MPS of 390 µm (Diet 1) or 511 μm (Diet 2). Diet 1 or 2 were randomly assigned within pig Groups 1 or 2 and 3 or 4. The digestive enzyme activities of duodenal fluid, particle size of intestinal digesta and feces, as well as nutrient digestibility, were determined for each pig as the experiment unit. The MPS of duodenal digesta (181 vs. 287 µm, p < 0.01), ileal digesta (253 vs. 331 µm, p < 0.01), and feces (195 vs. 293 µm, p < 0.01) was significantly reduced for pigs fed Diet 1 vs. Diet 2, respectively. Compared with Diet 2, Diet 1 significantly reduced the proportion of particles above 0.5 mm, but significantly increased the proportion of particles between 0.072 and 0.5 mm (p < 0.01) in digesta and feces (p < 0.01). Diet 1 significantly increased solubles percentage (<0.072 mm) in duodenal digesta (p < 0.05) but did not affect solubles percentage in ileal digesta and feces. The MPS of diet did not affect the activities of amylase, trypsin, and chymotrypsin in the duodenal fluid and the apparent total tract digestibility (ATTD) of dry matter, gross energy, crude protein, ether extract, neutral detergent fiber (NDF) and acid detergent fiber (ADF) in pigs offered Diet 1 compared to Diet 2. The in vitro digestible energy (IVDE) (3706 vs. 3641 kcal/kg; p = 0.03) was greater for Corn 1 vs. Corn 2. However, no significant difference was observed in IVDE (3574 vs. 3561 kcal/kg; p = 0.47) for Diet 1 vs. Diet 2. In conclusion, the particle size of digesta and feces was dependent on the dietary particle size. However, the digestive enzyme activities of duodenal fluid and ATTD of energy and nutrients were not affected by reducing dietary MPS from 511 to 390 µm.

Coated sodium butyrate supplementation to a reduced nutrient diet enhanced the performance and positively impacted villus height and faecal and digesta bacterial composition in weaner pigs

A study was conducted to evaluate the effects of dietary supplementation of different levels of coated sodium butyrate (CSB) on growth performance, nutrient digestibility, faecal and intestinal coliform and lactic acid bacteria counts, gas emission and small intestinal villi length in weaning pigs. A total of 160 weaning pigs [(Landrace x Yorkshire) x Duroc, 28 days old] with an average initial body weight (BW) of 7.04 ± 1.15 kg were randomly allotted to 4 treatments (8 replicates per treatment with 5 pigs; 3 barrows and 2 gilts per replicate pen) according to their initial body weight for a 6-week trial in three phases. Treatments consisted of corn-soybean meal-based diet with slight reduction in lactose (approximately 5 %) as well as sodium content relative to National Research Council (NRC, 2012) requirement as the control (CON) and CON diet supplemented with low dose CSB (0.5 g/kg feed), medium dose CSB (1.5 g/kg feed for first 3 weeks followed by reduction to 0.75 g/kg feed thereafter) and high dose CSB (3 g/kg feed for first 3 weeks followed by reduction to1.5 g/kg feed thereafter). As a result of this study, the supplementation of increasing levels of CSB increased final BW (linear, P = 0.04, quadratic P = 0.02 effects) at day 42, linearly increased average daily gain (ADG) and gain: feed (G:F) ratio (P = 0.05, P = 0.01 respectively) during day 8−21. The ADG showed trends (P = 0.073) in increment and a significant linear increase in G:F (P < 0.05) during days 1−21 were observed with the increase in the supplemental levels of CSB. The overall ADG was linearly increased (P < 0.05). The co-efficient of apparent total tract digestibility (ATTD) of dry matter (DM) was higher (linear effect, P = 0.001; quadratic effect, P = 0.01) with the increase in the levels of CSB during day 21 but not during day 42. A trend in reduction of coliform counts (linear effect, P = 0.08, quadratic effect, P = 0.07) was observed during day 21. The lactic acid bacteria count was increased (linear and quadratic effects, P = 0.01) in pigs fed diet supplemented with increasing levels of CSB at day 42. The coliform counts were reduced (linear and quadratic effects, P < 0.05) in the digesta obtained from duodenum of pigs fed high; medium and low doses of CSB supplemented diet. The supplementation of low, medium and high doses of CSB showed reduction (linear effect, P < 0.001, quadratic effect, P = 0.01) in fecal H2S concentration. The inclusion of increasing dose of CSB in the diet increased (linear effect, P < 0.05) the villi length of duodenum. In addition, linear (P < 0.05) and quadratic (P < 0.05) responses in villi length were observed in jejunum and ileum from pigs fed graded levels of CSB. Taken together, low or medium levels of CSB supplementation can have a beneficial role in enhancing performance, and digestibility of weaning pigs.

Utility Evaluation of Porcine Enteroids as PDCoV Infection Model in vitro.

Porcine deltacoronavirus (PDCoV) is a novel emerging enteric coronavirus found in pigs. Intestinal enteroids, which partially recreate the structure and function of intestinal villi-crypts, have many physiological similarities to the intestinal tissues in vivo. Enteroids exhibit advantages in studying the interactions between intestines and enteric pathogens. To create a novel infection model for PDCoV, we developed an in vitro system to generate porcine intestinal enteroids from crypts of duodenum, jejunum, and ileum of pigs. Enterocytes, enteroendocrine cells, Paneth cells, stem cells, proliferating cells, and goblet cells were found in the differentiated enteroids. Replication of PDCoV was detected in the cultured enteroids by immunofluorescence and quantitative RT-PCR. Double immunofluorescence labeling demonstrated that PDCoV was present in Sox9-positive intestinal cells and Villin1-positive enterocytes. There were multiple cellular responses shown as changes of transcription of genes related to mucosal immunity, antiviral genes, and marker genes of stem cells and other cells in the enteroids infected with PDCoV. We conclude that the 2-D enteroids derived from porcine jejunum can be used as an in vitro multicellular model for the investigation of pathogenesis and host immune responses to porcine enteric pathogens, such as PDCoV.

Dietary protein-bound or free amino acids differently affect intestinal morphology, gene expression of amino acid transporters, and serum amino acids of pigs exposed to heat stress.

Pigs exposed to heat stress (HS) increase body temperature in which can damage the intestinal epithelia and affect the absorption and availability of amino acids (AA). Protein digestion and metabolism further increase body temperature. An experiment was conducted with six pairs of pigs (of 47.3 ± 1.3 kg initial body weight) exposed to natural HS to assess the effect of substituting dietary protein-bound AA by free AA on morphology and gene expression of intestinal epithelial and serum concentration (SC) of free AA. Treatments were: high protein, 21.9% crude protein (CP) diet (HShp) and low protein, 13.5% CP diet supplemented with crystalline Lys, Thr, Met, Trp, His, Ile, Leu, Phe, and Val (HSaa). The HShp diet met or exceeded all AA requirements. The HSaa diet was formulated on the basis of ideal protein. Pigs were fed the same amount at 0700 and 1900 hours during the 21-d study. Blood samples were collected at 1700 hours (2.0 h before the evening meal), 2030 hours, and 2130 hours (1.5 and 2.5 h after the evening meal). At the end, all pigs were sacrificed to collect intestinal mucosa and a 5-cm section from each segment of the small intestine from each pig. Villi measures, expression of AA transporters (y+L and B0) in mucosa, and SC of AA were analyzed. Ambient temperature fluctuated daily from 24.5 to 42.6 °C. Weight gain and G.F were not affected by dietary treatment. Villi height tended to be larger (P ≤ 0.10) and the villi height:crypt depth ratio was higher in duodenum and jejunum of pigs fed the HSaa diet (P < 0.05). Gene expression of transporter y+L in jejunum tended to be lower (P < 0.10) and transporter B0 in the ileum was lower (P < 0.05) in HSaa pigs. Preprandial (1700 hours) SC of Arg, His, Ile, Leu, Thr, Trp, and Val was higher (P < 0.05), and Phe tended to be higher (P < 0.10) in HShp pigs. At 2030 hours (1.5 h postprandial), serum Lys, Met, and Thr were higher in the HSaa pigs (P < 0.05). At 2130 hours (2.5 h), Arg, His, Ile, Phe, and Trp were lower (P < 0.05); Met was higher (P < 0.05); and Lys tended to be higher (P < 0.10) in HSaa pigs. In conclusion, feeding HS pigs with low protein diets supplemented with free AA reduces the damage of the intestinal epithelia and seems to improve its absorption capacity, in comparison with HS pigs fed diets containing solely protein-bound AA. This information is useful to formulate diets that correct the reduced AA consumption associated with the decreased voluntary feed intake of pigs under HS.

Oregano Feed Supplementation Affects Glycoconjugates Production in Swine Gut.

Simple SummaryThe European ban towards antibiotics is increasing the number of studies on the effects of feed additives, such as plant extracts, in order to enhance the health and welfare status of domestic animals intended for human consumption. Origanum vulgare possesses multiple pharmacological characteristics and its antioxidant and antibacterial properties are particularly interesting. Besides, a recent study aimed at evaluating the effects of oregano aqueous extract supplementation in poultry nutrition gave encouraging results regarding the secretion of glycoconjugates in the gut which increases tissue hydration and protects the intestinal mucosa from pathogenic bacteria, viruses, and parasites. Therefore, we investigated the effects of oregano feed supplementation on antioxidant and defense ability of pig gut. Our results showed that there was improved production of glycoconjugates in the duodenum and colon of pigs fed with supplementation of oregano aqueous extract, enhancing protection of the mucosa of these sections of the intestine. Also, we observed an enhanced antioxidant action in the two examined gut tract samples of the group supplemented with oregano. Findings can be used in further research to identify ways to improve endogenous defense ability with to reduce antibiotic use and prevent antimicrobial resistance.This study evaluated the effects of adding oregano aqueous extract (OAE) to the diet of pig slaughtered at finisher stage. Study was performed to identify glycoconjugates and evaluate the oxidative stress levels in the duodenum and colon intestinal tracts. Glycohistochemistry was performed by staining with Periodic acid–Schiff (PAS), Alcian blue (AB) pH 2.5, AB-PAS, AB pH 1, AB pH 0.5, low iron diamine, and high iron diamine. Serial sections were pre-treated with sialidase V before staining with AB pH 2.5 (Sial-AB) preceded or not by saponification. To study oxidative stress, an immunohistochemical analysis was applied to investigate the presence of the oxidative stress target molecule Bcl-2 Associate X protein (BAX). Findings show that oregano aqueous extract supplementation improves the production of the secretion glycoconjugates involved in direct and indirect defense, thus enhancing the protection of the pig intestinal mucosa. Moreover, the reduced BAX protein immunostaining observed in both duodenum and colon of swine of the oregano-supplemented group respect to that observed in the control group suggests an enhanced antioxidant action by oregano adding. Findings could be useful for other studies aiming to reduce antibiotic use and prevent antimicrobial resistance.

Fructooligosaccharides improve growth performance and intestinal epithelium function in weaned pigs exposed to enterotoxigenic Escherichia coli.

To explore the protective effect of Fructooligosaccharides (FOS) against Enterotoxigenic Escherichia coli (ETEC)-induced inflammation and intestinal injury, twenty-four weaned pigs were randomly assigned into three groups: (1) non-challenge (CON, fed with basal diet), (2) ETEC-challenge (ECON, fed with basal diet), and (3) ETEC challenge + FOS treatment (EFOS, fed with basal diet plus 2.5 g kg-1 FOS). On day 19, the CON group was orally infused with sterilized culture while pigs in the ECON group and EFOS group were orally infused with ETEC (2.5 × 1011 colony-forming units). After 3 days, pigs were slaughtered for sample collection. We showed that ETEC challenge significantly reduced average daily gain (ADG); however, FOS improved the ADG (P < 0.05), apparent digestibility of crude protein (CP), gross energy (GE), and ash and reduced the diarrhea incidence (P < 0.05). FOS reduced plasma concentrations of IL-1β and TNF-α and down-regulated (P < 0.05) the mRNA expression of IL-6 and TNF-α in the jejunum and ileum as well as IL-1β and TNF-α in the duodenum. The concentrations of plasma immunoglobulin A (IgA), immunoglobulin M (IgM) and secreted IgA (SIgA) in the jejunum (P < 0.05) were elevated. Interestingly, FOS elevated the villus height in the duodenum, and elevated the ratio of villus height to crypt depth in the duodenum and ileum in the EFOS group pigs (P < 0.05). Moreover, FOS increased lactase activity in the duodenum and ileum (P < 0.05). The activities of sucrase and alkaline phosphatase (AKP) were higher in the EFOS group than in the ECON group (P < 0.05). Importantly, FOS up-regulated the expressions of critical genes in intestinal epithelium function such as zonula occludens-1 (ZO-1), L-type amino acid transporter-1 (LAT1), and cationic amino acid transporter-1 (CAT1) in the duodenum and the expressions of ZO-1 and glucose transporter-2 (GLUT2) in the jejunum (P < 0.05). FOS also up-regulated the expressions of occludin, fatty acid transporter-4 (FATP4), sodium glucose transport protein 1 (SGLT1), and GLUT2 in the ileum (P < 0.05). FOS significantly increased the concentrations of acetic acid, propionic acid and butyric acid in the cecal digesta. Additionally, FOS reduced the populations of Escherichia coli, but elevated the populations of Bacillus and Bifidobacterium in the caecal digesta (P < 0.05). These results suggested that FOS could improve the growth performance and intestinal health in weaned pigs upon ETEC challenge, which was associated with suppressed inflammatory responses and improved intestinal epithelium functions and microbiota.

Effect of fibre sources on performance, serum parameters, intestinal morphology, digestive enzyme activities and microbiota in weaned pigs

ABSTRACTThis study was conducted to evaluate the effects of wheat bran (WB) as insoluble fibre source, and sugar beet pulp (SBP) as soluble fibre source, on performance, serum parameters and intestinal health in weaned pigs. A total of 90 weaned pigs (BW: 7.33 ± 1.24 kg) were randomly assigned to three dietary treatments: (1) a control diet (CON) based on corn and soybean meal; (2) CON + 6% WB; (3) CON + 6% SBP. Each treatment had five replicate pens with six pigs per pen. The experimental period was divided into two phases (d 0 to 14 and d 14 to 28). Pigs in group WB tended to have greater avarage daily gain than those in group SBP. Compared with CON, SBP reduced (p < 0.05) the apparent total tract digestibility (ATTD) of dry matter, organic matter (OM), gross energy and neutral detergent fibre on d 14 and 28, while WB decreased (p < 0.05) the ATTD of OM on d 28. Pigs in group SBP had higher (p < 0.05) glucagon-like peptide 1 concentration than the other groups on d 14 and 28. The villus height to crypt depth (V:C) ratio of duodenum and jejunum in pigs fed diet WB were greater (p < 0.05) than in group SBP. The WB increased (p < 0.05) the V:C ratio of ileum compared with CON or SBP. Compared with SBP, WB increased (p < 0.05) the sucrase activity in the duodenum. Moreover, pigs in WB had higher (p < 0.05) activities of maltase and sucrase in the jejunum compared with CON or SBP. The abundances of Ruminococcaceae and Prevotellaceae were increased (p < 0.05) in WB, while the Lachnospiraceae abundance was increased (p < 0.05) in SBP. WB increased (p < 0.05) concentrations of acetate, butyrate and total short-chain fatty acids (SCFA), while SBP increased (p < 0.05) concentrations of acetate and total SCFA when compared with CON. In conclusion, WB was beneficial to performance in weaned pigs by improving morphology, enzyme activities and microbiota when compared with SBP, highlighting that effects of fibre depends on the fibre sources.

Validation of an Ex Vivo Permeation Method for the Intestinal Permeability of Different BCS Drugs and Its Correlation with Caco-2 In Vitro Experiments.

The absorption study of drugs through different biological membranes constitutes an essential step in the development of new pharmaceutical dosage forms. Concerning orally administered forms, methods based on monolayer cell culture of Caco-2 (Caucasian colon adenocarcinoma) have been developed to emulate intestinal mucosa in permeability studies. Although it is widely accepted, it has disadvantages, such as high costs or high technical complexity, and limitations related to the simplified structure of the monolayer or the class of molecules that can be permeated according to the transport mechanisms. The aim of this work was to develop a new ex vivo methodology which allows the evaluation of the intestinal apparent permeability coefficient (Papp) while using fewer resources and to assess the correlation with Caco-2. To this end, pig (Sus scrofa) duodenum segments were mounted in Franz diffusion cells and used to permeate four different drugs: ketorolac tromethamine (Kt), melatonin (Mel), hydrochlorothiazide (Htz), and furosemide (Fur). No statistically significant differences (p > 0.05) were observed corelating Papp values from Franz diffusion cells and Caco-2 cell experiments for Kt, Htz, and Fur. However, there were statistically significant differences (p < 0.05) correlating Papp values and Mel. The difference is explained by the role of Mel in the duodenal epithelial paracellular permeability reduction. Ex vivo permeation may be an equivalent method to Caco-2 for drugs that do not produce intestinal membrane phenomena that could affect absorption.

Enhanced GII.4 human norovirus infection in gnotobiotic pigs transplanted with a human gut microbiota.

The role of commensal microbiota in enteric viral infections has been explored extensively, but the interaction between human gut microbiota (HGM) and human norovirus (HuNoV) is poorly understood. In this study, we established an HGM-Transplanted gnotobiotic (Gn) pig model of HuNoV infection and disease, using an infant stool as HGM transplant and a HuNoV GII.4/2006b strain for virus inoculation. Compared to germ-free Gn pigs, HuNoV inoculation in HGMT Gn pigs resulted in increased HuNoV shedding, characterized by significantly higher shedding titres on post inoculation day (PID) 3, 4, 6, 8 and 9, and significantly longer mean duration of virus shedding. In addition, virus titres were significantly higher in duodenum and distal ileum of HGMT Gn pigs on PID10, while comparable and transient HuNoV viremia was detected in both groups. 16S rRNA gene sequencing demonstrated that HuNoV infection dramatically altered intestinal microbiota in HGMT Gn pigs at the phylum (Proteobacteria, Firmicutes and Bacteroidetes) and genus (Enterococcus, Bifidobacterium, Clostridium, Ruminococcus, Anaerococcus, Bacteroides and Lactobacillus) levels. In summary, enhanced GII.4 HuNoV infection was observed in the presence of HGM, and host microbiota was susceptible to disruption upon HuNoV infection.

L-phenylalanine Increased Gut Hormone Secretion through Calcium-Sensing Receptor in the Porcine Duodenum.

Simple SummaryThe proper modulation of feed intake not only meets the nutrient requirement for the maximum growth rate in pigs, but also avoids feed waste. A complete understanding of how dietary factors affect the appetite in pigs will provide a strategy to modulate the feed intake of pigs. L-phenylalanine (L-Phe) has been demonstrated to induce satiety through stimulating anorectic hormone secretion in rodents. However, whether L-Phe has similar effects in pigs is unknown. Here, we aimed to investigate how L-Phe affects gut hormone secretion, along with insight into the underlying mechanism in porcine duodenum by using an in vitro perfusion system. Results showed that 80 mM L-Phe triggered glucose-dependent insulinotropic peptide (GIP) and cholecystokinin (CCK) release, and also upregulated calcium-sensing receptor (CaSR) and its downstream molecules, such as protein kinase C (PKC) and inositol 1,4,5-triphosphate receptor (IP3R) expression. However, these effects were attenuated by treatment with a CaSR antagonist. Our findings show that CaSR participates in Phe-induced hormone secretion in pig duodenum, indicating that CaSR may be a potential target in the food intake regulation of pigs.Luminal amino acids have a pivotal role in gut hormone secretion, and thereby modulate food intake and energy metabolism. However, the mechanisms by which amino acids exert this effect remains unknown. The purpose of this research was to investigate the response of L-phenylalanine (L-Phe) to gut hormone secretion and its underlying mechanisms by perfusing the pig duodenum. Eighty mM L-Phe and extracellular Ca2+ stimulated cholecystokinin (CCK) and glucose-dependent insulinotropic peptide (GIP) release, and upregulated the mRNA expression of the calcium-sensing receptor (CaSR), CCK, and GIP. Western blotting results showed that L-Phe also elevated the protein levels of CaSR, the inositol 1,4,5-triphosphate receptor (IP3R), and protein kinase C (PKC). However, the CaSR inhibitor NPS 2143 reduced the mRNA expression of CaSR, CCK, and GIP, and the secretion of CCK and GIP, as well as the protein level of CaSR, IP3R, and PKC. These results indicated that Phe stimulated gut secretion through a CaSR-mediated pathway and its downstream signaling molecules, PKC and IP3R.

Effects of hot melt extrusion processed nano-iron on growth performance, blood composition, and iron bioavailability in weanling pigs.

This study was conducted to investigate the effects of hot melt extrusion (HME) nano-iron as an alternative for the common ferrous sulfate on iron (Fe) bioavailability, growth performance, nutrient digestibility, intestinal morphology, and intestinal microbiota of weanling pigs. A total of 200 piglets (Landrace × Yorkshire × Duroc) were randomly allotted to seven treatments on the basis of initial body weight (BW) and sex. Treatments were the INO100 (100 ppm Fe as FeSO4), HME-Fe levels (50, 75, and 100 ppm nano-Fe as FeSO4). ORG100 (100 ppm Fe as iron methionine). In phase 1, the HME50 pigs showed the lowest Fe content in feed and feces. Plasma Fe concentration was increased in HME100 and ORG100 pigs. In phase 2, there were significantly lower concentration of Fe in feed and feces of HME50 pigs (p < 0.01). A lower Fe concentration in the plasma and liver were observed in HME50 pigs compared with HME100 pigs. Concentration of red blood cell (RBC) was the lowest (p < 0.01) for HME50 pigs. During phase 2, the HME100, HME75, and ORG100 pigs showed a higher RBC and hemoglobin values compared with HME50 pigs. Digestibility of gross energy (GE) and crude protein (CP) were significantly higher in HME100 pigs compared with HME50 pigs. There was an increased (p < 0.01) villus height in the duodenum and jejunum of HME100 pigs compared with HME50 pigs. It is concluded that dietary Fe does not improve growth performance of weanling pigs; however, increasing the dietary iron concentration in weanling piglets increased the RBC and hemoglobin. In addition, the potential ability of HME to be used at a lower level (HME75) was observed.

A comparison of dynamic distributions of intestinal microbiota between Large White and Chinese Shanxi Black pigs.

Intestinal microbiota has been widely recognized to influence on their hosts with respect to digestion and absorption of nutrients, but little is known about the structure and composition of microbial communities at different growth periods of hosts as yet. In this case, 16S rRNA gene amplicon sequencing was applied to decode the microbiota architecture in four distinct intestinal compartments (duodenum, jejunum, ileum, and cecum) of both Large White pigs and Chinese Shanxi Black pigs at the weaning, nursery, and fast-growth developmental stages. In our study, the intestinal ecosystems were dynamically changing and influenced by host maturity and diets at different development stages. Species phylogenetically affiliated to phyla Firmicutes, Proteobacteria, and Bacteroidetes were abundant in both pig breeds; at the genus level, microbial communities were dominated by Prevotella, followed by Acinetobacter and Lactobacillus. Further inspection revealed that Lactobacillus was identified to be positively associated with villus height, whereas Acinetobacter and Prevotella were prone to reside in deep crypts. Furthermore, intestinal microbiota in Shanxi Black pigs had more metabolic and less infectious functions than that in Large White pigs. In short, our data present here indicated that microbiota with longitudinal diversity and lower infection in Shanxi Black pigs might contribute to the relatively stronger adaptability in comparison with Large White pigs.

Sensing of L-Arginine by Gut-Expressed Calcium Sensing Receptor Stimulates Gut Satiety Hormones Cholecystokinin and Glucose-Dependent Insulinotropic Peptide Secretion in Pig Model.

Nutrients regulate the secretion of gut satiety hormones, which is related to the modulation of food intake and blood glucose levels. Calcium-sensing receptor (CaSR) is involved in regulating gut hormone secretion in response to l-amino acids and multivalent cations. Rodents are often used to investigate the effect of nutrients on these hormonal release. However, results obtained using rodent models are difficult to be applied in humans, we used pigs as a model in this study because their physiology is similar to that of humans. In this study, we investigated whether l-Arginine (l-Arg) could induce gut hormones cholecystokinin (CCK) and glucose-dependent insulinotropic peptide (GIP) secretion in the porcine duodenum and if so, whether CaSR mediated l-Arg-regulated gut satiety hormone secretion. Our data showed that treatment with 20 and 50 mM l-Arg induced CCK and GIP secretion compared with 0 mM l-Arg. However, treatment with d-Arg (an inactive isomer) failed to elicit this response. The potency of l-Arg to induce CCK and GIP secretion was enhanced in the presence of extracellular Ca2+ and CaSR agonist cinacalcet. However, the effect of Arg on CCK and GIP secretion was attenuated by blocking CaSR and its downstream signaling molecules adenylate cyclase (AC) and phospholipase C (PLC). Taken all together, pig duodenum provides an appropriate model to explore the effects of l-Arg on the secretion of the satiety-related gut hormones CCK and GIP and the role of CaSR in this effect. Further investigations are needed to verify the effect of l-Arg on food intake and blood glucose in human study. PRACTICAL APPLICATION: l-Arginine is able to modulate cholecystokinin and glucose-dependent insulinotropic peptide secretion through the CaSR in pig model, which has a potential role in regulating food intake and blood glucose levels.

Effect of Lactobacillus salivarius on growth performance, diarrhea incidence, fecal bacterial population and intestinal morphology of suckling pigs challenged with F4+ enterotoxigenic Escherichia coli.

ObjectiveGut health improvements were monitored with respect to growth performance, diarrhea incidence, fecal bacterial population and intestinal morphology of suckling pigs orally supplemented with live Lactobacillus salivarius (L. salivarius) oral suspensions and challenged with F4+ enterotoxigenic Escherichia coli (ETEC).MethodsTwo groups of newborn pigs from 18 multiparous sows were randomly designated as non-supplemented (control: n = 114 piglets) and L. salivarius supplemented groups (treatment: n = 87 piglets). Treatment pigs were orally administered with 2 mL of 109 colony-forming unit (CFU)/mL L. salivarius on days 1 to 3, then they were orally administered with 5 mL of 109 CFU/mL L. salivarius on days 4 to 10, while those in control group received an equal amount of phosphate buffered saline solution. On day 24 (2 weeks post supplementation), one pig per replicate of both groups was orally administered with 108 CFU/mL F4+ ETEC, then they were euthanized on day 29 of experiment.ResultsResults revealed that pigs in treatment group had a statistically significant increase in average daily gain, body weight and weight gain, and tended to lower diarrhea throughout the study. Numbers of Lactobacillus population in feces of treatment pigs were higher than control pigs, especially on day 10 of study. Numbers of total bacteria in intestinal contents of control pigs were also increased, but not Coliform and Lactobacillus populations. Histological examination revealed statistically significant improvements of villous height and villous/crypt ratio of duodenum, proximal jejunum and distal jejunum parts of treatment pigs compared with controls. Duodenal pH of treatment group was significantly decreased.ConclusionOral supplementation of live L. salivarius during the first 10 days of suckling pig promoted growth performance and gut health, reduced diarrhea incidence, increased fecal Lactobacillus populations and improved intestinal morphology.

Pyrosequencing-based analysis of the complex microbiota located in the gastrointestinal tracts of growing-finishing pigs

The commensal gut microbial communities play an important role in the health and production efficiency of growing-finishing pigs. This study aimed to analyse the composition and diversity of the microbiota in the gastrointestinal tract sections (stomach, duodenum, jejunum, ileum, caecum, colon and rectum) of growing-finishing pigs. This analysis was assessed using 454 pyrosequencing targeting the V3–V6 region of the 16S rRNA gene. Samples were collected from 20, healthy pigs aged 24 weeks and weighing 115.9 ± 5.4 kg. The dominant bacterial phyla in the various gastrointestinal tract sections were Firmicutes, Bacteroidetes, Proteobacteria and Actinobacteria. At the genus level, Prevotella, unclassified Lachnospiraceae, Ruminococcus, unclassified Ruminococcaceae and Oscillospira were more abundant in the large intestine than in the stomach and the small intestine. Unclassified Peptostreptococcaceae and Corynebacterium were more abundant in the small intestine than in the stomach and the large intestine. Shuttleworthia, unclassified Veillonellaceae and Mitsuokella were more abundant in the stomach than in the small and large intestines. At the species level, M. el.s.d.enii and M. multacida were predominant in the stomach. In addition, P. stercorea, P. copri, C. butyricum, R. flavefaciens and R. bromii were significantly more abundant in the large intestine than in the stomach and the small intestine. B. pseudolongum and B. thermacidophilum were significantly more abundant in the small intestine than in the stomach and the large intestine. Principal coordinates analysis showed that the overall composition of the pig gastrointestinal microbiota could be clustered into three groups: stomach, small intestine (duodenum, jejunum and ileum) and large intestine (caecum, colon and rectum). Venn diagrams illustrated the distribution of shared and specific operational taxonomic units among the various gastrointestinal tract sections.

A flagellin-adjuvanted PED subunit vaccine improved protective efficiency against PEDV variant challenge in pigs

PEDV variants, causing severe diarrhea in neonatal suckling piglets with a mortality rate up to 100%, have being epidemic since late 2010 in china. To meet the pressing need of safe and cost-efficient PED maternal vaccines against PEDV variant, we vaccinated growing piglets with a flagellin-adjuvanted PED vaccine rSF-COE-3D by injection at Houhai acupoint. The vaccination not only enhanced the antibody responses of serum IgG/IgA, mucosal IgA and serum neutralizing antibody, but also improved the production of IFN-γ and IL-4. Moreover, rSF-COE-3D could provide a better protection against the challenge of a high pathogenic PEDV variant, with less diarrhea pigs, less pigs with detectable PEDV shed, lower rank values of feces and less apparent lesions and inflammation in duodenum of the PEDV infected pigs. The improved protective efficiency of rSF-COE-3D compared with COE was mostly benefited from the enhanced production of serum IgA, mucosal IgA in feces and serum neutralizing antibody titers. Taken together, our data suggest that rSF-COE-3D would be a novel efficient PED vaccine.

Effects of dietary energy and lipase levels on nutrient digestibility, digestive physiology and noxious gas emission in weaning pigs.

ObjectiveThis study was conducted to evaluate the effect of dietary energy and lipase supplementation on growth performance, nutrient digestibility, serum profiles, intestinal morphology, small intestinal digestive enzyme activities, biochemical index of intestinal development and noxious gas emission in weaning pigs.MethodsA total of 240 weaning pigs ([Yorkshire×Landrace]×Duroc) with an average body weight (BW) of 7.3±0.12 kg were used in this 28-d experiment. Weaning pigs were randomly allocated to 4 dietary treatments in a 2×2 factorial arrangement with 2 levels of energy (net energy = 2,470 kcal/kg for low energy diet and 2,545 kcal/kg for basal diet) and 2 levels of lipase (0 and 1.5 U/g of lipase) according to BW and sex. There were 6 replications (pens) per treatment and 10 pigs per pen (5 barrows and 5 gilts).ResultsWeaning pigs fed the low energy diet had lower (p<0.05) gain-to-feed ratio (G:F) throughout the experiment, apparent digestibility of dry matter, nitrogen, ether extract, and gross energy during d 0 to 14, average daily gain during d 15 to 28, lipase activity in duodenum and ileum and protein/DNA in jejunum (p<0.05), respectively. Lipase supplementation had no effect on growth performance but affected apparent nutrient digestibility (p<0.05) on d 14 and enhanced lipase activity in the duodenum and ileum and protease activity in duodenum and jejunum of pigs (p<0.05) fed the low energy diet. Lipase reduced serum low-density lipoprotein cholesterol (LDL-C) and triglyceride (TG), NH3 production (p<0.05) from the feces.ConclusionThe low energy diet decreased G:F throughout the experiment and nutrient digestibility during d 0 to 14 as well as lipase activity in duodenum and ileum. Lipase supplementation increased nutrient digestibility during d 0 to 14 and exerted beneficial effects on lipase activity in duodenum and ileum as well as protease activity in duodenum and jejunum, while reduced serum LDL-C, TG and fecal NH3.

Probiotic supplementation affects the glycan composition of mucins secreted by Brunner’s glands of the pig duodenum

The effect of a dietary probiotic blend on the carbohydrate composition of mucins secreted by the Brunner’s glands in the duodenum of growing-finishing pigs was investigated by means of conventional (periodic acid-Schiff, Alcian Blue pH 2.5, high iron diamine staining) and lectin (15 lectins) histochemistry. Pigs were assigned to two dietary treatments: a control basal diet without the probiotic blend (No-Pro) and a test diet that included the probiotic blend (Pro). Duodenal tissue fragments were fixed in 4% phosphate-buffered-saline-buffered paraformaldehyde, dehydrated through a graded alcohol series, and embedded in paraffin wax. The secretory cells of the Brunner’s glands from No-Pro pigs primarily produced neutral glycoproteins and a small amount of acidic non-sulphated mucins. This glycan pattern was opposite that of the Brunner’s glands from Pro animals. A comparison of lectin-binding profiles of the secretory cells of Brunner’s glands in these two groups showed that in Pro pigs, there was (i) a decrease in N-linked glycans containing α1,2-linked fucose (Con A, UEA I); (ii) a loss of complex types of N-glycans (PHA-L, PHA-E) terminating with lactosamine (RCA120), α1,6- and α1,3-linked fucose (LTA), and α-galactose (GSA I-B4), as well as of O-glycans with terminal Galβ1,3GalNAc (PNA); and (iii) an increase in O-glycans containing GalNAc HPA. No-Pro and Pro samples showed no change in the expression of α2,6 sialoglycans and terminal GlcNAc residues and no affinity for MAL II, DBA, and SBA. These results indicate that probiotic supplementation affects the glycan composition of mucins produced in the Brunner’s glands of growing-finishing pigs. These changes could effectively act on the gastrointestinal function and health status of these animals because the probiotic blend induced higher growth performance and meat quality in the test probiotic group than it did in the control basal diet group (Tufarelli et al., 2017).

The influence of probiotic fodder additives on the morphofunctional state of duodenum pigs

The article is dedicated to the study of the effect of a class of microorganisms and substances of microbial and other origin that are used in feeding pigs to achieve maximum growth in animal mass, as well as therapeutic purposes. Study of the effect of biologically active additives on the state of the pig's digestive system. The article considers the rational nutrition that increase the productivity and reproductive capacity of animals, and also prevent the emergence of profound disturbances of all metabolic processes which leads to a decrease in resistance to productivity, a clinically pronounced disease of adult animals and young animals. The authors pay attention to the morphometric parameters, ultrastructure and content of nucleic acids in the wall of the duodenum of pigs by forage feeding with the addition of probiotic fodder additive «Probion-forte» in dose of 1 g/kg of fodder. Research were conducted on 28 day pigs, breed «Large White». It was formed two groups of piglets per 30 heads; piglets were fed with standard mixed fodders; piglets from the first group were received standard mixed fodder with the addition of probiotic fodder additive «Probion-forte» in dose 1 g/kg of fodder for 42 days. After the slaughter of piglets, pieces of duodenum for histological, histochemical and ultrastructural studies were selected. Statistical significance of differences was determined by Student's t test , assuming 5% estimate error. It was shown that feeding with forage within 42 days from the addition of probiotic fodder additive «Probion-forte» at a dose of 1 g/kg, villus height is increase, crypt depth and a number of plasma cells in the lamina propria of mucosa of the duodenum, which helps the digestive process and increase the area of nutrient absorption in the intestines. The number of plasma cells are increased in the lamina propria of mucosa and testify immunomodulatory effect of fodder additives. Ultra structural alteration of microvilli and changes in the nuclei of duodenal enterocytes of piglets of the first group indicates a more pronounced acfunctional activity of enterocytes and thereby increases the activity of parietal digestion in the intestine. As a conclusion, the task by definition performed of the effective dose of probiotic in feed for pig it can be considered completed and we can state the probiotics can be incorporated, as a alternative to antibiotics and increase the weight gain of animals and also to increase the level of immune resistance of the organism.