Crypt Depth Ratio Research Articles

Effects of dietary xylanase supplementation on growth performance, intestinal health, and immune response of nursery pigs fed diets with reduced metabolizable energy.

This study aimed to investigate the effects of xylanase on growth performance and intestinal health of nursery pigs fed diets with reduced metabolizable energy (ME). One hundred ninety-two pigs at 8.7 kg ± 0.7 body weight (BW) after 7 d of weaning were allotted in a randomized complete block design with initial BW and sex as blocks. Eight dietary treatments consisted of 5 ME levels (3,400, 3,375, 3,350, 3,325, and 3,300 kcal ME/kg) below the NRC (2012) requirement and 4 levels of xylanase (0, 1,200, 2,400, and 3,600 XU/kg) to a diet with 3,300 kcal ME/kg. All pigs received their respective treatments for 35 d in 2 phases, pre-starter (14 d) and starter (21 d). On day 35, eight pigs in 3,400 kcal/kg (CON), 3,300 kcal/kg (LE), and 3,300 kcal/kg + 3,600 XU xylanase/kg (LEX) were euthanized to collect jejunal tissues and digesta for the evaluation of mucosa-associated microbiota, intestinal immune response, oxidative stress status, intestinal morphology, crypt cell proliferation, and digesta viscosity as well as ileal digesta to measure apparent ileal digestibility. Data were analyzed using the MIXED procedure on SAS 9.4. The LE increased (P < 0.05) jejunal digesta viscosity, tended to have decreased (P = 0.053) relative abundance of Prevotella, and tended to increase (P = 0.055) Lactobacillus. The LE also increased (P < 0.05) the concentration of protein carbonyl whereas malondialdehyde, villus height (VH), villus height to crypt depth ratio (VH:CD), apparent ileal digestibility (AID) of nutrients, and finally average daily feed intake were decreased (P < 0.05). The LE did not affect average daily gain (ADG). The LEX decreased (P < 0.05) digesta viscosity, increased (P < 0.05) the relative abundance of Prevotella, decreased (P < 0.05) Helicobacter, decreased (P < 0.05) the concentration of protein carbonyl, tended to increase (P = 0.065) VH, and decreased (P < 0.05) VH:CD and crypt cell proliferation. Moreover, LEX increased (P < 0.05) the AID of dry matter and gross energy and tended to increase (P = 0.099; P = 0.076) AID of crude protein, and ether extract. The LEX did not affect ADG but did tend to decrease (P = 0.070) fecal score during the starter phase. Overall, reducing ME negatively affected intestinal health parameters and nutrient digestibility without affecting growth. Supplementation of xylanase mitigated some of the negative effects observed by ME reduction on intestinal health and digestibility of nutrients without affecting growth.

Effects of β-mannanase supplementation on intestinal health and growth of nursery pigs.

Two experiments were conducted using 120 pigs to test the hypothesis that supplementation of β-mannanase could reduce digesta viscosity, enhance nutrient digestion, and improve intestinal health and growth of nursery pigs. In experiment 1, 48 crossbred barrows were randomly allotted to four treatments with increasing levels of β-mannanase at 0, 200, 400, and 600 U/kg in feeds. All pigs were euthanized on day 12 to collect jejunal digesta to measure digesta viscosity and ileal digesta to measure apparent ileal digestibility (AID) of dry matter (DM), gross energy (GE), neutral detergent fiber (NDF), and acid detergent fiber (ADF). In experiment 2, 72 nursery pigs were randomly allotted to three treatments with increasing levels of β-mannanase at 0, 400, and 600 U/kg in feeds. Plasma collected on day 9 was used to measure tumor necrosis factor-α (TNF-α), immunoglobulin G (IgG), malondialdehyde (MDA), and protein carbonyl (PC). All pigs were euthanized on day 10 to collect duodenal and jejunal tissues to evaluate the production of TNF-α, IL-6, and MDA, morphology, crypt cell proliferation, and expression of tight junction proteins in the jejunum. Data were analyzed using the MIXED procedure for polynomial contrasts and the NLMIXED procedure for broken-line analysis of SAS. In experiment 1, β-mannanase supplementation tended to have quadratic effects on digesta viscosity (P = 0.085) and AID of GE (P = 0.093) in the pigs. In experiment 2, jejunal digesta viscosity of the pigs was reduced (P < 0.05) when β-mannanase was supplemented at 360 U/kg of feed. β-Mannanase supplementation linearly reduced (P < 0.05) TNF-α, IgG, MDA, and PC in the duodenum, and TNF-α, IgG, and MDA in the jejunum of the pigs. β-Mannanase supplementation linearly increased (P < 0.05) villus height to crypt depth ratio and crypt cell proliferation in the jejunum. β-Mannanase supplementation tended to linearly improve (P = 0.083) expression of zonula occludens-1 in the jejunum. In conclusion, supplementation of β-mannanase at 360 U/kg reduced the digesta viscosity and up to 600 U/kg positively affected intestinal health and growth of pigs by reducing inflammation and oxidative stress whilst enhancing structure and barrier function in the jejunum.

Feather meal processing methods impact the production parameters, blood biochemical indices, gut function, and hepatic enzyme activity in broilers.

This study investigated the effects of feather meal (FM) processing methods on production parameters, blood biochemical indices, intestinal morphology, digestive and hepatic enzyme activities, and gastrointestinal tract pH and microflora of broilers. A total of 480-d-old male broilers were used for 42 d in a completely randomized design with eight treatments and five replicates (12 chicks/replicate). Treatments were 1) a control diet (without FM), 2) a diet containing 4% raw FM (RFM), 3) a diet containing 4% processed FM (PFM) by autoclave (Au-PFM), 4) a diet containing 4% fermented FM (FFM) by Bacillus licheniformis (Bl-FFM), 5) a diet containing 4% FFM by Bacillus subtilis (Bs-FFM), 6) a diet containing 4% FFM by Aspergillus niger (An-FFM), 7) a diet containing 4% FFM by B. licheniformis + B. subtilis + A. niger (Co-FFM), and 8) a diet containing 4% PFM by an enzyme (En-PFM). Results showed that in the FFMs the contents of ash, ether extract, total volatile nitrogen, and amino acids including Lys, Met, Thr, Trp, His, Leu, Gly, Ile, Phe, and Tyr increased (P < 0.05), while crude fiber, crude protein, and dry matter content decreased (P < 0.05). Compared with the control, the Co-FFM diet had no significant differences (P > 0.05) in total body weight gain (2,827 vs. 2,791g/chick), total feed intake (5,018 vs. 4,991g/chick), European production efficiency factor (375 vs. 377), European Broiler Index (371 vs. 371), and feed conversion ratio (1.77 vs. 1.78g/g). Feeding FFM decreased (P < 0.05) serum total cholesterol (1.46-fold), triglyceride (1.61-fold), very low-density lipoprotein cholesterol (1.61-fold), and low-density lipoprotein cholesterol (2.27-fold) compared to the control. Also, FFM increased (P < 0.05) villus height (1,045 to 1,351, 661 to 854, and 523 to 620 μm), and villus height to crypt depth ratio (6.15 to 8.45, 4.55 to 7.04, and 4.27 to 5.45), in the duodenum, jejunum, and ileum, respectively, compared to the control. Compared to the control, the Co-FFM diet increased (P < 0.05) protease (34, 39, and 45 %) in the pancreas, duodenum, and jejunum, as well as amylase (73, and 97 %) activities in the duodenum, and jejunum, respectively. Diets containing FFM reduced (P < 0.05) pH in the crop, gizzard, and ileum, and decreased (P < 0.05) Escherichia coli (6.12 to 5.70) count in ileum compared to the control. The Co-FFM diet increased (P < 0.05) lactic acid bacteria count in crop (6.77 to 7.50) and ileum (6.94 to 7.73), also decreased (P < 0.05) coliforms (6.31 to 5.75) count in ileum compared to the control. In conclusion, FM fermentation, particularly Co-FFM, improves the nutritional value of FM, converting it into a decent source of dietary protein for broilers.

Microbioma intestinal e morfometria de codornas alimentadas com dietas contendo óleos essenciais

ABSTRACT: One of the greatest challenges in poultry production is maintaining intestinal mucosal barrier integrity and gut microbiota balance. Safe alternative antimicrobials that can regulate the microbial community through animal feed have been the subject of research in poultry production. This study evaluated the effect of Mentha piperita and Melaleuca alternifolia essential oils (EOs) on the gut microbiome and morphometry of broiler quails under normal feeding conditions. The gut microbiome was studied using a completely randomized design consisting of 4 treatments, namely control, bacitracin zinc, and the Eos M. piperita and M. alternifolia, with 8 repetitions and 7 quails per treatment, totaling 224 quails from 1 to 42 days old. The intestinal contents of the slaughtered quails were collected to evaluate the gut microbiome profile of their digestive tract. Gut morphometry was analyzed using a completely randomized factorial design, with four experimental rations for three intestinal sections (4x3) and five replications. The variables studied were villus surface area and height, crypt depth, villus height to cryptdepth ratio (VH:CD), villus-crypt ratio (V:C), villus width to height ratio (VW:H), and height of the intestinal epithelium and musculature. M. alternifolia (50 mg/kg of feed) in the diet of broiler quails improved gut morphometry, similar to the results obtained with bacitracin zinc. This EO also altered the gut microbiome of quails and reduced pathogenic bacterial diversity.

Effects of fat sources on liver characteristics and intestinal morphometry in an early-life animal model.

Motivation for the study. There is little evidence on whether the consumption of fat sources containing different proportions of fatty acids has an effect on the characteristics of the liver and small intestine at an early age. Main findings. We found that the intake of fat sources containing unsaturated fatty acids contributes to maintaining the characteristics of both organs; whereas, consumption of sources containing saturated fatty acids favors inflammation in the liver and small intestine. Implications. The consumption of quinoa oil constitutes an alternative to protect these noble organs in an animal model at an early age. We aimed to determine the effect of the consumption of three sources of fatty acids on the relative weight, macroscopic and microscopic characteristics of the liver, and intestinal morphometry in an early-life animal model. Seventy-six randomly distributed chicks received one of the diets (T1: 97.0% basal diet (BD) + 3.0% inert material, T2: 97.0% BD + 3.0% partially hydrogenated vegetable shortening, T3: 97.0% BD + 3.0% quinoa oil, and T4: 97.0% BD + 3.0% fish oil) until the seventh day of life; samples were then extracted in order to be analyzed. We found that the animals that consumed quinoa oil (T3) or fish oil (T4) had favorable results associated to lower liver weight and better absorption of nutrients at intestinal level due to higher values in the hair length and crypt depth ratio, in comparison to partially hydrogenated vegetable shortening (T2). In conclusion, quinoa oil constitutes a healthy option for consumption and an alternative source to fish oil.

Protective influence of supplementary betaine against heat stress by regulating intestinal oxidative status and microbiota composition in broiler chickens.

To assess the impact of supplementing betaine (BT) under heat stress (HS) conditions on broiler performance and intestinal health from 21 to 42days of age, a total of 150 male Ross 308 broilers were indiscriminately allotted to 3 treatments with 10 replications of 5 birds each. The control (CON) group was given a basal ration and accommodated at a thermoneutral condition (22 ± 1°C), whereas the HS and HS + BT groups were raised under cyclic HS (33 ± 1°C for 8h and 22 ± 1°C for 16h per day) and received the basal ration without or with 1000mg/kg BT, respectively. The HS reduced average daily gain (ADG); average daily feed intake; villus height (VH); VH to crypt depth (CD) ratio (VCR); activities of trypsin, lipase, glutathione peroxidase (GPX), and catalase; and enumeration of Lactobacillus and Bifidobacterium (P < 0.05) and augmented feed conversion ratio (FCR), CD, malondialdehyde (MDA) accumulation, and enumeration of Escherichia coli, Clostridium, and coliforms (P < 0.05). Conversely, BT supplementation heightened ADG, VH, VCR, trypsin activity, GPX activity, and populations of Lactobacillus and Bifidobacterium (P < 0.05) and lowered FCR, MDA accumulation, and Clostridium population (P < 0.05). Furthermore, the FCR value, trypsin and GPX activities, MDA content, and Bifidobacterium and Clostridium populations in the HS + BT group were nearly equivalent to those in the CON group. To conclude, feeding BT under HS conditions could improve broiler performance through improving intestinal health by specifically mitigating oxidative damage and enhancing the colonization of beneficial bacteria.

Growth performance, serum parameters, inflammatory responses, intestinal morphology and microbiota of weaned piglets fed 18% crude protein diets with different ratios of standardized ileal digestible isoleucine to lysine

The present study was to explore the Ile requirement of piglets fed 18% crude protein (CP) diets. Two hundred and fifty 28-day-old Duroc × Landrace × Yorkshire piglets (8.37 ± 1.92 kg) were randomly divided into 5 dietary treatments (10 piglets per replicate, 5 barrows and 5 gilts per replicate) with 45%, 50%, 55%, 60%, 65% standardized ileal digestible (SID) Ile-to-Lys ratios, and the SID Lys was formulated to 1.19%. The experimental design consisted of two phases (d 1 to 14 and d 15 to 28). Results showed that average daily gain (ADG) had a tendency to quadratically increase as the SID Ile-to-Lys ratio increased (P = 0.09), and the optimum SID Ile-to-Lys ratios required to maximize ADG were 48.33% and 54.63% for broken-line linear model and quadratic polynomial model, respectively. Different SID Ile-to-Lys ratios had no significant effects on average daily feed intake and gain-to-feed ratio. Dry matter (P < 0.01), CP (P = 0.01), ether extract (P = 0.04), gross energy (P < 0.01) and organic matter (P < 0.01) digestibility increased quadratically. Serum total cholesterol levels decreased linearly (P = 0.01) and quadratically (P < 0.01); aspartate aminotransferase (P < 0.01), interleukin-1β (P = 0.01), and tumor necrosis factor-α (P < 0.01) levels decreased quadratically; immunoglobulin G (P = 0.03) and immunoglobulin M (P = 0.01) concentrations increased quadratically. Serum Ser levels decreased linearly (P < 0.01) and quadratically (P = 0.01); Glu (P = 0.02), Arg (P = 0.05), and Thr (P = 0.03) levels decreased quadratically; Gly (P < 0.01) and Leu (P = 0.01) levels decreased linearly; Ile (P < 0.01) concentration increased linearly. Duodenal villus height (P < 0.01) and villus height to crypt depth ratio (P < 0.01) increased quadratically. The deficiency or excess of Ile decreased short chain fatty acid-producing bacteria abundance and increased pathogenic bacteria abundance. Overall, taking ADG as the effect index, the optimum SID Ile-to-Lys ratios of piglets offered 18% CP diets were 48.33% and 54.63% based on two different statistical models, respectively, and the deficiency or excess of lle negatively affected piglet growth rates and health status.

Methionine sources and genotype affect embryonic intestinal development, antioxidants, tight junctions, and growth-related gene expression in chickens

Methionine (Met) is an essential and first limiting amino acid in the poultry diet that plays a significant role in chicken embryonic development and growth. The present study examined the effect of in ovo injection of DL-Met and L-Met sources and genotypes on chicken embryonic-intestinal development and health. Fertilized eggs of the two genotypes, TETRA-SL layer hybrid (TSL) — commercial layer hybrid and Hungarian Partridge colored hen breed (HPC) — a native genotype, were randomly distributed into four treatments for each genotype. The treatment groups include the following: 1) control non-injected eggs (NoIn); 2) saline-injected (SaIn); 3) DL-Met injected (DLM); and 4) L-Met injected (LM). The in ovo injection was carried out on 17.5 d of embryonic development; after hatching, eight chicks per group were sacrificed, and the jejunum was extracted for analysis. The results showed that both DLM and LM groups had enhanced intestinal development as evidenced by increased villus width, villus height, and villus area (P < 0.05) compared to the control. The DLM group had significantly reduced crypt depth, glutathione (GSH) content, glutathione S-transferase 3 alpha (GST3), occludin (OCLN) gene expression and increased villus height to crypt depth ratio in the TSL genotype than the LM group (P < 0.05). The HPC genotype has overexpressed insulin-like growth factor 1 (IGF1) gene, tricellulin (MD2), occludin (OCLN), superoxide dismutase 1 (SOD1), and GST3 genes than the TSL genotype (P < 0.05). In conclusion, these findings showed that in ovo injection of Met enhanced intestinal development, and function, with genotypes responding differently under normal conditions. Genotypes also influenced the expression of intestinal antioxidants, tight junction, and growth-related genes.

Effect of green tea (Camellia sinensis L.) as antimicrobial agent on growth performance and ileal histomorphology of broiler chickens

ABSTRACT We investigated the impact of green tea powder (GTP) as an alternative to antibiotics on growth performance and gut health in broilers. A total of 340 one-day-old broilers were assigned to four dietary treatments as control (free of antibiotic), antibiotic, GTP 0.5 g/kg (GTP-1) and GTP 1 g/kg (GTP-2). Overall, the antibiotic group demonstrated significantly (P < 0.05) higher feed intake, weight gain, lower feed conversion ratio and better dressing percentage compared to the GTP groups, and the control group had the lowest values. GTP-1 showed the highest (P < 0.05) villus height (VH), followed by the antibiotic group and GTP-2, while the control group had the lowest villus height. The control group displayed a higher (P < 0.05) crypt depth, followed by GTP-2, while the antibiotic group and GTP-1 had significantly (P < 0.05) lower crypt depth. The villus height to crypt depth ratio was significantly higher in GTP-1, followed by similar values in the antibiotic group and GTP-2, while the control group had the lowest ratio (P < 0.05). From the results of the present study, we concluded that GTP at the level of 0.5 g/kg of feed is an excellent dose vis-à-vis antibiotic in broiler for optimum growth performance and gut health.

Effect of Phytobiotics and Antibiotic on Growth Performance, Intestinal Morphology and Nutrients Transporters Expression of Broiler Chickens

This study investigated the effect of Phytobiotics (PB) and Antibiotic Growth Promoter (AGP) on performance, intestinal morphology and nutrients transporters mRNA gene expression of broiler chickens. One hundred eighty unsexed one-day-old Ross strains with an average initial body weight (BW) of 45 ± 0.5 g were randomly assigned into three treatments with six replications of 10 chicks in each pen. (1) Control group (CON), basal diets only; (2) antibiotic group, Antibiotic Growth promoter (AGP), basal diet supplemented with 5 mg/kg tetramycin; (3) Phytobiotics (PB), basal diet supplemented with 0.5g/kg, phytobiotics the research lasted for 42 days. Average Daily Feed Intake (ADFI), Average Daily Gain (ADG) and Feed Conversion Ratio (FCR) were calculated weekly, and intestinal morphology of Villus Height (VH), Crypt Depth (CD), and Villus Height to crypt depth ratio (VH: CD) from the duodenum, jejunum, and ileum, nutrients transporters qRT-PCR mRNA expression of GLUT1, SGLT4, GLUT5, PEPT2, CAT1, and LAT1 were evaluated at day 42. Chicks fed PB had significantly (P=0.0413) higher ADG at 1-42 days, PB and AGP significantly had lower FCR (P=0.0001) and (P=0.0001) at 22-42 d and 1-42 days, respectively, in duodenum the villus height was significantly (P=0.0001) longer in PB fed chickens and the lowest was recorded in CON, while the CD and VH: CD were significantly longer in AGP and PB fed chickens, in jejunum VH and CD were significant (P=0.0001) longer in PB and AGP than the CON fed chickens, in ileum PB fed birds had significant higher VH and CD than the AGP and CON fed groups, in duodenum and ileum GLUT1, SGLT4 and GLUT5 expression were highly expressed in AGP and PB fed chickens similar results was obtained in jejunum and ileum by PEPT2 and CAT1. These findings suggest that supplementing phytobiotics in broiler diets improves growth performance and intestinal morphology and up-regulated the expression of nutrient transporters' genes. However, the underlying detailed biological mechanisms and dose standardisation for inclusion in broiler diets need to be further studied.

A Composite Morphometric Duodenal Biopsy Mucosal Scale for Celiac Disease Encompassing Both Morphology and Inflammation

Villus height to crypt depth ratio (Vh:Cd) and intraepithelial lymphocytes (IEL) are key measures of histology of the small intestine in celiac disease. Although the field of celiac disease has advanced, there remains no broadly accepted measure of mucosal injury. We assessed whether a composite Vh:Cd and IEL scale (VCIEL) can improve accuracy and statistical precision for assessing histology, compared with individual measures. The formulation of the VCIEL composite histologic scale was based on combining the Vh:Cd and IEL measurements for individual patients with equal weighting, by converting each scale to a fraction of their standard deviation and summing the results. The VCIEL formula was applied to several clinical trials and the results for Vh:Cd and IEL were compared with those for VCIEL with regards to clinical significance (effect size) and statistical significance. For the ALV003-1021 trial, we observed an effect size and P value (analysis of covariance) of 1.37 and 0.038 for ΔVh:Cd, 1.17 and 0.005 for ΔIEL, and 1.86 and 0.004 for ΔVCIEL. For the similar gluten-challenge IMGX003-NCCIH-1721 trial, the corresponding results were 0.76 and 0.057 for ΔVh:Cd, 0.98 and 0.018 for ΔIEL, and 1.14 and 0.007 for ΔVCIEL. Similar improvements with the use of VCIEL over individual Vh:Cd and IEL measures were observed for other studies, including a nontherapeutic gluten challenge study. The composite VCIEL scale combining Vh:Cd and IEL values seems to improve accuracy and statistical precision compared with either component alone.

PSVI-13 Dietary Milk Fat Globule Membrane Improves Neonatal Piglet Intestinal Architecture and Enhances Mucosal Diamine Oxidase Activity Following Lipopolysaccharide Challenge

Abstract Early life stressors alter the trajectory of gut development and impair nutrient utilization, barrier function, and animal performance throughout life. Gastrointestinal (GI) maladies increase morbidity and mortality in neonates, and dietary nutrients are essential in supporting GI function. Although maternal milk is the gold-standard for yielding developmental benefits to offspring, genetic selection in the modern sow has resulted in large litter size and milk yield potentially insufficient to achieve the high biological growth potential in sow-reared piglets. Research into milk quality has led to the identification of many bioactive nutrients important in neonatal development. Components of the milk fat globule membrane (MFGM) contribute immensely to neonatal nutrition and gut health. However, mechanisms of actions are still being defined. The objective is to define the effect of MFGM supplementation on neonatal piglet intestinal health. We hypothesize dietary MFGM will enhance piglet intestinal architecture and tight junction (TJ) protein abundance, attenuating systemic lipopolysaccharide (LPS)-induced intestinal barrier disruption. One-day-old piglets (n = 32) were assigned to soy (CON) or MFGM phospholipid supplemented diet (0.75% wt/wt) ± LPS (100 µg/kg body weight) challenge in a 2 × 2 factorial design (n = 8/diet/challenge). Piglets were fed for 21 d and 8 animals/diet received saline or LPS injections 4 h before euthanasia. Piglet weights and feed intake were recorded daily to calculate performance. Intestinal tissues were collected to assess villus height (VH) and crypt depth (CD) by hematoxylin and eosin staining. Two biomarkers of intestinal integrity, intestinal fatty acid-binding protein (I-FABP) and diamine oxidase (DAO), were measured in serum or tissue. Protein abundance of claudin-1 and occludin in ileal and colonic mucosa were detected by Western blot. Data were analyzed by 2 × 2 factorial arrangement of traetments using the PROC MIXED. Dietary MFGM had no adverse impact on growth performance or TJ proteins (P &amp;gt; 0.05). Dietary MFGM increased VH (CON: 348.8 and MFGM: 427.0 ± 26.79 µm) and decreased CD (CON: 166.3 and MFGM: 147.4 ± 8.42 µm) in ileum, thereby increasing the VH: CD ratio (CON: 2.2; MFGM: 3.1 ± 0.13; P &amp;lt; 0.05). Further, LPS treatment increased serum I-FABP by 4.4-fold and decreased jejunal mucosal DAO by 0.5-fold compared with saline treatment (P &amp;lt; 0.05). Additionally, there was a 93.1 % increase in colon DAO activity with MFGM diet compared with soy-fed pigs, regardless of LPS challenge (P &amp;lt; 0.05). Dietary MFGM improved neonatal piglet ileum architecture and increased colon DAO activity, potentially attenuating the intestinal barrier disruption following LPS challenge.

PSXIII-7 Effect of Dietary Supplementation of Lactobacillus Rhamnosus on Blood Profiles, Intestinal Morphology, Immune Responses, and Gene Expression of Tight Junction Proteins and Cytokines of Weaned Pigs

Abstract Dietary probiotics were used in the animal feeds because of the beneficial effect of improving gut health and immune responses as one of the potential alternatives to antibiotics. This study was conducted to evaluate the effect of dietary supplementation of Lactobacillus rhamnosus (ILDONG Bioscience Co. Ltd., Pyeongtaek, Republic of Korea) on blood profiles, small intestinal morphology, immune responses, and gene expression of tight junction proteins and inflammatory cytokines in ileum of weaned pigs. A total of 40 weaned pigs [6.21 ± 0.55 kg of initial body weight (BW); 28-day old] were allotted to 2 dietary treatments (4 pigs/pen; 5 replicates/treatment) in a randomized complete block design (block: initial BW) and fed for 4 weeks. Dietary treatments were 1) a commercial basal nursery diet (CON), and 2) CON + 0.02% Lactobacillus rhamnosus IDCC 3201 (PRO). The blood samples were collected from randomly selected 1 pig per pen on day 1, 7, 14, and 28 for blood profiles and immune responses. On the last day of experiment, the pigs were euthanized to collect duodenal, jejunal, and ileal segments for intestinal morphology and gene expression analysis. Intestinal morphology was measured villus height, width, area, crypt depth, villus height to crypt depth ratio, and number of goblet cells by hematoxylin and eosin staining method. The blood profiles were measured by using an automated hematology analyzer. The serum samples were used to measure concentrations of cytokines and cortisol by using porcine immunoassay ELISA kits. The gene expression of tight junction proteins and cytokines were measured by qRT-PCR. Data were analyzed by PROC GLM procedure of SAS. Experimental unit was a pen. Statistical model for blood profiles, immune responses, and intestinal morphology included dietary treatment as a main effect and initial BW as a covariate. The t-test was used for ileal gene expression. Pigs fed PRO had lower (P &amp;lt; 0.05) platelet level, crypt depth in duodenum and ileum, serum tumor necrosis factor-α (TNF-α), and interleukin-6 than those fed CON. The pigs fed PRO downregulated (P &amp;lt; 0.05) TNF-α and interleukin-8 genes compared with those fed CON, but upregulated (P &amp;lt; 0.05) claudin-1 and claudin-2 genes. In conclusion, probiotic Lactobacillus rhamnosus supplementation in nursery diet improved gut health through modulating blood profiles and immune responses, enhancing intestinal morphology, and regulating tight junction protein genes expression in ileum of weaned pigs.

273 Effects of Xylanase Supplementation in Diets with Reduced Metabolizable Energy and Supplemental Fat on Intestinal Health, Mucosa Associated Microbiota and Growth Performance of Nursery Pigs

Abstract This study aimed to investigate the effects of reducing metabolizable energy (ME) below the requirement by reducing supplemental fat on intestinal health and growth performance of nursery pigs. One hundred and ninety-two newly weaned pigs at 7.3 ± 0.6 kg BW were allotted in a RCBD with sex and initial BW as blocks. Pigs were assigned to one of 8 dietary treatments with varying ME levels (3,400, 3,375, 3,350, 3,325, and 3,300 kcal/kg ME) by reducing supplemental fat and increasing xylanase (X0: 0, X40: 1,200, X80: 2,400 and X120: 3,600 XU/kg of feed) in ME deficient diets by 100 kcals/kg. Pigs were fed diets for 35 d in 2 phases (14 d and 21 d, respectively) and growth performance was measured at the end of each phase. Phase 2 diet had titanium dioxide (0.4%) as indigestible external marker to measure ileal digestibility of nutrients. At d 35, a pig representing the median BW within each pen from 3 dietary treatments (ME at 3,400 and 3,300 kcal/kg ME with or without xylanase) were euthanized for sampling jejunal mucosa, jejunal tissue, and ileal digesta to analyze intestinal health parameters, mucosa associated microbiota, and apparent ileal digestibility (AID) of nutrients. Data were analyzed using Mixed procedure of SAS 9.4. Overall, reducing ME level had no effect on growth performance of nursery pigs. However, reducing ME by 100 kcal/kg ME below the requirement increased (P &amp;lt; 0.05) digesta viscosity (1.7 to 2.2 cP) and protein carbonyl (0.18 to 0.40 nmol/mg), tended to change the relative abundance of Prevotellaceae (P = 0.062, 49 to 23%) and Lactobacillaceae (P = 0.086, 7 to 19%), reduced (P &amp;lt; 0.05) villus height (467 to 433 µm) and villus height to crypt depth ratio (1.80 to 1.64) in the jejunum, and reduced (P &amp;lt; 0.05) AID of DM (59.5 to 47.3%), gross energy (59.1 to 43.0%), crude protein (63.4 to 44.6%), and ether extract (67.7 to 55.2%). Xylanase supplementation decreased (P &amp;lt; 0.05) digesta viscosity (2.2 to 1.82 cP) and protein carbonyl (0.40 to 0.17 nmol/mg), increased the relative abundance of Prevotellaceae (20.22 to 61.56%), increased villus height (433 to 462 µm) and villus height to crypt depth ratio (1.64 to 1.99) in the jejunum, and increased (P &amp;lt; 0.05) AID of DM (47.3 to 58.7%), and gross energy (42.9 to 56.0%). In conclusion, reducing ME (100 kcal/kg below the requirement) by reducing supplemental fat (by 2% unit) impaired intestinal health, reducing ability of the pigs to digest feeds whereas xylanase supplementation appeared to mediate some of the negative effects seen by energy reduction by improving intestinal morphology, reducing oxidative damage products, and diversity of the mucosa-associated microbiota.

PSXIII-8 Effects of Dietary Lactobacillus Reuteri on Immune Responses, Blood Profiles, Intestinal Morphology, and Ileal Gene Expression of Weaned Pigs

Abstract Weaning period is critical for gut health and immunity of pigs due to negative effect of weaning stress. Probiotics can be a potential alternative to antibiotics. However, there is limited information on potential mechanism of gut health and immunity of dietary Lactobacillus reuteri. This study was conducted to investigate the effects of dietary Lactobacillus reuteri on immune responses, blood profiles, intestinal morphology, and ileal gene expression of weaned pigs. A total of 48 weaned pigs [average initial body weight (BW) = 6.51 ± 0.87 kg; 21 days old] were allotted to 2 dietary treatments in a randomized complete block design (block: initial BW and sex). Dietary treatments had 6 replicate pens with 4 pigs per pen and fed for 4 weeks. Dietary treatments consisted of a basal weaner diet based on corn-soybean meal (CON) and CON supplemented with 0.02% Lactobacillus reuteri IDCC 3701 (LR). The LR was obtained from a commercial company (ILDONG Bioscience Co., Ltd., Pyeongtaek, Republic of Korea). Blood samples were collected from randomly selected one pig per pen on day 1, 7, 14, and 28. Serum was obtained from the blood sample to measure immune responses. On the last day of the experiment, the pigs were euthanized to collect small intestinal segments for intestinal morphology. Ileal mucosa were collected from ileal tissue for gene expression analysis. Measurements were serum cortisol, tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), interleukin-1β (IL-1β), and interleukin-6 (IL-6) using ELISA kits, blood profiles using an automated hematology analyzer, intestinal morphology using hematoxylin and eosin method, and ileal gene expression of tight junction proteins and inflammatory cytokines using qRT-PCR. Data were analyzed by using PROC GLM of SAS. Experimental unit was a pen. Statistical model for immune responses, blood profiles, and intestinal morphology included dietary treatments as a main effect and initial BW as a covariate. The t-test was used for the comparison of ileal gene expression. Pigs fed LR had drecreased (P &amp;lt; 0.05) serum TNF-α and IL-6 on day 7 and serum TGF-β1 on day 14 than those fed CON. In addition, pigs fed LR had greater (P &amp;lt; 0.05) hemoglobin concentration on day 7 than those fed CON, but had reduced (P &amp;lt; 0.05) hematocrit on day 14. Pigs fed LR had greater (P &amp;lt; 0.05) villus height to crypt depth ratio in duodenum than those fed CON. The LR upregulated (P &amp;lt; 0.05) claudin-1 gene compared with CON, but downregulated (P &amp;lt; 0.05) interleukin-8 and TNF-α genes. In conclusion, addition of dietary Lactobacillus reuteri in weaner diets improved intestinal morphology, modulated blood profiles and immune responses, and gene expression of inflammatory cytokines.

258 Effects of Dietary Supplementation of Myristic Acid on Intestinal Health and Growth Performance of Nursery Pigs

Abstract The objective of this study was to investigate the effects of myristic acid on intestinal health and growth performance of nursery pigs. Thirty-six newly weaned pigs (6.6 ± 0.4 kg BW) were assigned to 3 treatments (n = 12) in a randomized complete block design with initial BW and sex as blocks. Pigs were fed for 35 d in 3 phases (10, 10, and 15 d respectively). Treatments were 1) NC: basal diet; 2) PC: NC + 0.25% bacitracin methylene disalicylate (antibiotic; bacitracin: 276 g/t feed) in all phases; and 3) MA: NC + myristic acid at 0.075% in phases 1 and 2 and 0.045% in phase 3. Diets contained nutrients meeting the requirement. Growth performance and fecal score were measured for each phase. Pigs were euthanized to collect jejunal mucosa, jejunal tissues, and ileal digesta. Data were analyzed using preplanned contrasts (NC vs. PC and NC vs. MA) in the Proc MIXED procedure of SAS. Compared with the NC, the PC improved (P &amp;lt; 0.05) ADG in phase 1 (43 to 101 g/d) and ADFI in phase 2 (458 to 578 g/d), whereas the MA improved (P &amp;lt; 0.05) ADG (541 to 668 g/d) and ADFI (801 to 996 g/d) in phase 3. Overall, the PC tended to improve ADG (353 to 419 g/d; P = 0.063), and ADFI (515 to 606 g/d; P = 0.051), whereas the MA improved (P &amp;lt; 0.05) ADG (353 to 431 g/d) and ADFI (515 to 623 g/d) compared with the NC group. The PC decreased (P &amp;lt; 0.05) fecal score (3.17 to 3.03) in phase 2 compared with the NC. The PC tended to decrease IL8 (P = 0.053; 0.53 to 0.37 ng/mg of protein) and protein carbonyl (P = 0.075; 2.65 to 1.66 nmol/mg of protein) in jejunal mucosa compared with the NC. The PC increased (P &amp;lt; 0.05) the villus height to crypt depth ratio (2.61 to 3.02) compared with that of NC. The MA tended to decrease IgG (P = 0.051; 2.44 to 1.67 µg/mg of protein) and IL8 (P = 0.090; 0.53 to 0.39 ng/mg of protein) in jejunal mucosa compared with that of the NC. The MA tended to increase relative abundance of Bacteroidetes (P = 0.075; 3.8 to 13.9) at phylum level, Prevotellaceae (P = 0.073; 3.7 to 13.8) at family level, and Prevotella (P = 0.085; 4.2 to 15.4) at genus level compared with the NC. In this study, it was shown that both bacitracin and myristic supplementation could improve the intestinal health and growth performance of nursery pigs. Effects of bacitracin on intestinal health and growth performance was rather immediate whereas the effects of myristic acid were obtained after a 3-week feeding.

PSIV-27 Effect of Different Ratios of Phytogenic Feed Additives in Weaned Pigs Challenged with a Pathogenic Escherichia Coli

Abstract This study was conducted to investigate the effects of supplementing different ratios of phytogenic feed additive (PFA) to weaned pigs challenged with pathogenic Escherichia coli (E. coli) on growth performance, nutrient digestibility, intestinal barrier integrity, and immune response, and to determine the optimal mixing ratio for PWD prevention. A total of 48 four-week-old weaned pigs with initial body weight of 8.01 ± 0.39 kg were placed in individual metabolic cages, and then randomly assigned to eight treatment groups. The eight treatments were as follows: a basal diet without E. coli challenge (negative control, NC), a basal diet with E. coli challenge (positive control, PC), PC with supplementing 0.1% mixture of 20 % bitter citrus extract (BCE), 10% microencapsulated blend of thymol and carvacrol (MEO), and 70% excipient (T1), PC with supplementing 0.1% mixture of 10% MEO, 20% premixture of grape seed and grape marc extract, green tea, and hops (PGE), and 60% excipient (T2), PC with supplementing 0.1% mixture of 10% BCE, 10% MEO, 10% PGE, and 70% excipient (T3), PC with supplementing 0.1% mixture of 20% BCE, 20% MEO, and 60% excipient (T4), PC with supplementing 0.1 % mixture of 20% MEO, 20% PGE, and 60% excipient (T5), and PC with supplementing 0.1 % mixture of 10% BCE, 20% MEO, 10% PGE, and 60% excipient (T6; Table 1). The experiments were progressed in 16 days, including 5 days before and 11 days after the first E. coli challenge (d 0). In the E. coli challenge treatments, all pigs were orally inoculated by dividing a total of 10 mL of E. coli F 18 for 3 consecutive days from d 0 post-inoculation (PI). Compared with the PC group, the PFA2 and PFA6 groups significantly increased (P &amp;lt; 0.05) feed efficiency and decreased (P &amp;lt; 0.05; Table 2) diarrhea during the entire period (Table 3). At d 11 PI, the PFA6 group significantly improved (P &amp;lt; 0.05) gross energy digestibility compared with the PFA1 group (Table 4). The PFA6 group significantly decreased (P &amp;lt; 0.05) tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) in serum (Table 5) and increased (P &amp;lt; 0.05) the villus height to crypt depth ratio (VH:CD; Table 6; Figure 1). The PFA2 significantly decreased (P &amp;lt; 0.05) the relative protein expression of calprotectin in the ileum (Table 7). In conclusion, improvements in growth performance, diarrhea reduction, and immunity enhancement are demonstrated when 10% BCE, 20% MEO, 10% PGE, and 60% excipient are mixed.

PSXIII-4 Effects of Probiotic Lactobacillus Rhamnosus on Intestinal Morphology, Blood Profiles, Immune Responses, and Ileal Gene Expression of Weaned Pigs

Abstract Probiotics are considered as a potential non-antibiotic feed additive that may improve intestinal health of pigs. However, there is limited information on potential mechanism of probiotic Lactobacillus rhamnosus. The study was carried out to evaluate effects of probiotic Lactobacillus rhamnosus on intestinal morphology, blood profiles, immune responses, and ileal gene expression of weaned pigs. In a randomized complete block design [block: initial body weight (BW)], a total of 100 weaned pigs (7.99 ± 0.85 kg of average initial BW; 28 days of age) were assigned to two dietary treatments (5 pigs/pen; 10 replicates/treatment). Dietary treatments were fed for 4 weeks: 1) a commercial basal nursery diet (CON), and 2) CON + 0.02% probiotic Lactobacillus rhamnosus IDCC 3201 (PRO). Probiotic Lactobacillus rhamnosus was obtained from a commercial company (ILDONG Bioscience Co., Ltd., Pyeongtaek, Republic of Korea). Blood was collected from randomly selected one pig per pen on day 1, 7, 14, and 28 for blood profiles and immune responses. On the final day of study, pigs were euthanized to collect each part of small intestine for measuring intestinal morphology and ileal gene expression. Measurements were 1) villus height, width, and area, crypt depth, villus height to crypt depth ratio (VH:CD), and number of goblet cells by the hematoxylin and eosin staining method, 2) number of white blood cells (WBC), red blood cells (RBC), and hematocrit (HCT) by an automated hematology analyzer calibrated for porcine blood, 3) immunoglobulin G (IgG), IgA, tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), interleukin-10 (IL-10), and C-reactive protein in serum by the ELISA kits, and 4) ileal gene expression of tight junction-related proteins and inflammatory cytokines by qRT-PCR. Data were analyzed using PROC GLM of SAS. Experimental unit was the pen. Statistical model for intestinal morphology, blood profiles, and immune responses included dietary treatment as a main effect and initial BW as a covariate. The t-test was used for ileal gene expression. The PRO increased number of goblet cells in duodenum (P &amp;lt; 0.05) and VH:CD in ileum (P &amp;lt; 0.10) compared with CON. Pigs fed PRO had decreased HCT on day 7 (P &amp;lt; 0.10), WBC on day 28 (P &amp;lt; 0.05), and serum IgA concentrations on day 28 (P &amp;lt; 0.05) than those fed CON, but RBC on day 7 and 14 were greater (P &amp;lt; 0.05). The PRO upregulated (P &amp;lt; 0.05) claudin-3, occludin, tight junction protein-1 genes in ileum compared with CON, but TNF-α, interferon-α, IL-1α, IL-1β, IL-6, and IL-8 genes were downregulated (P &amp;lt; 0.05). In conclusion, addition of probiotic Lactobacillus rhamnosus in the nursery diet improved gut health via enhancing intestinal morphology and upregulating ileal tight junction-related protein genes, and modified blood parameters and ileal inflammatory cytokine genes.

Physiological changes in broiler chickens subjected to dietary ajwain (trachyspermum ammi l.) essential oil in encapsulated and conventional forms within a wheat-based diet

The aim of this study was to investigate the effects of different diet types, forms, and contents of ajwain essential oil (AEO) on various physiological characteristics of broiler chickens, including cell-mediated immunity responses, intestinal morphology, and microflora. A total of 1500 one-day-old male broiler chickens were allocated to different treatments based on a 2 × 3 × 2 factorial arrangement, considering diet types (corn and corn-wheat), contents of AEO (0, 150, and 300 mg/kg of diet), and forms of AEO (conventional and encapsulated). The results indicated that the broiler chickens fed the diet containing 150 ppm EO demonstrated reduced skin thickness in response to a 2,4-dinitrochlorobenzene challenge, 24 h after injection, compared to those receiving a diet without EO (P < 0.05). Increasing the EO content led to an increase in the villous height to crypt depth ratio in the jejunum of broiler chickens receiving 300 ppm EO (P < 0.05). Moreover, there was a slight improvement in the villous height to crypt depth ratio in the jejunum of broiler chickens fed the corn-wheat diet (P = 0.07). Broiler chickens fed the 300 ppm EO showed a lower total bacterial population compared to those fed the 150 ppm EO (P < 0.05). Finally, the use of EO at a content of 150 ppm improved cellular immune response, while EO at a content of 300 ppm improved the morphology and overall population of intestinal bacteria. Furthermore, the inclusion of wheat-corn diets exhibited enhanced morphological characteristics of the intestines. However, the forms of AEO did not exert any significant influence on the physiological traits.

136 Awardee Talk: Effects of Monoglycerides on Intestinal Morphology and Immune Responses of Weanling Pigs Experimentally Infected with a Pathogenic Escherichia Coli

Abstract This study aimed to investigate the effects of monoglycerides on intestinal morphology and immune responses of weanling pigs experimentally infected with Escherichia coli (E. coli) F18. Weaned pigs [n = 60; body weight (BW) = 6.49 ± 0.74 kg; around 21 d old] were individually housed and fed with one of four diets (15 replicates/diet): 1) control diet (CON); 2) CON+0.3% monoglycerides; 3) CON+3000 mg/kg ZnO; and 4) CON+50 mg/kg antibiotics (carbadox). The experiment lasted 28 days with 7 days before and 21 days after first inoculation (d 0). All piglets were orally challenged with E. coli F18 (1010 CFU/3 mL) for 3 consecutive days. Serum samples were collected on d 2, 5, and 21 post-inoculation (PI) to measure the concentration of tumor necrosis factor (TNF) -α, C-reactive protein, and haptoglobin. Duodenum, jejunum, and ileum segments were collected on d 5 PI (6 pigs/treatment) and on 21 PI (9 pigs/treatment) to measure intestinal morphology. Jejunal and ileal mucosa were also collected to measure the mRNA expression of immune responses related genes. Data were analyzed by ANOVA using PROC MIXED of SAS with randomized complete block design. Supplementation of high dose ZnO tended (P = 0.056) to reduce serum haptoglobin on d 0, reduced (P &amp;lt; 0.05) serum haptoglobin on d 2 and 5 PI, and reduced (P &amp;lt; 0.05) serum C-reactive protein on d 21 PI. Pigs fed with monoglycerides had less (P &amp;lt; 0.05) serum haptoglobin on d 5 PI, compared with CON. Pigs supplemented with high dose ZnO had more (P &amp;lt; 0.05) goblet cell numbers per villus, greater (P &amp;lt; 0.05) villi area and villi height, and greater (P &amp;lt; 0.05) villi height:crypt depth ratio in duodenum than pigs in other treatments on d 5 PI. Supplementation of monoglycerides, high dose of ZnO, or antibiotics reduced (P &amp;lt; 0.05) ileal crypt depth compared with CON on d 5 PI. Pigs in high dose ZnO group tended (P = 0.064) to have the largest villi height:crypth depth ratio in the ileum, followed by pigs in monoglycerides and antibiotics groups on 5 PI. Consistently, pigs fed with high dose ZnO expressed less (P &amp;lt; 0.05) TNF α, IL6, IL10, IL12, IL1A, IL1B, and PTGS2 in ileal mucosa, compared with other treatments on d 5 PI. However, no difference in the expression of listed genes was observed between pigs fed with monoglycerides vs. high dose of ZnO. Pigs supplemented with monoglycerides expressed least (P &amp;lt; 0.05) PTGS2 in ileal mucosa compared with other treatments on d 21 PI. In conclusion, dietary supplementation of 0.3% monoglycerides reduced intestinal and systemic inflammation of weaned pigs infected with E. coli F18, although the efficacy was not comparable to high dose ZnO.