Ileal Samples Research Articles

Determination of digestibility of different forages in dairy cows using indigestible NDF as marker

Four 4×4 Latin square experiments were conducted with four cannulated dairy cows. Eight forages were fed ad libitum or supplemented with concentrate. Digestibility and flow of neutral detergent fibre (NDF) were evaluated using chromic oxide as a marker. Additionally, duodenal and ileal flows were determined based on faecal flow of in situ estimated indigestible NDF (iNDF) as marker, where faecal flow of iNDF was determined using chromic oxide. No significant difference was found between intake of iNDF and faecal flow of iNDF calculated using chromic oxide as a marker, whereas ileal flow of iNDF accounted for 0.85±0.02 and duodenal flow of iNDF for 0.89±0.02 of faecal flow of iNDF, respectively. Using chromic oxide as a marker underestimates intestinal flows of fibre. Average iNDF:NDF ratios were 0.22, 0.39, 0.54, 0.40 and 0.54 in feed, rumen content, duodenal chyme, ileal chyme and faeces, respectively, indicating severe problems in obtaining representative ileal samples.

Treatment With a Novel Hemigramicidin-TEMPO Conjugate Prolongs Survival in a Rat Model of Lethal Hemorrhagic Shock

We sought to develop a therapeutic agent that would permit prolongation of survival in rats subjected to lethal hemorrhagic shock (HS), even in the absence of resuscitation with asanguinous fluids or blood. We synthesized a series of compounds that consist of the electron scavenger and superoxide dismutase mimic, 4-amino-2,2,6,6-tetramethylpiperidine-N-oxyl (4-NH2-TEMPO), conjugated to fragments and analogs of the membrane-active cyclopeptide antibiotic, gramicidin S. Using an in vivo assay, wherein isolated intestinal segments were loaded inside the lumen with various test compounds, we studied these compounds for their ability to prevent ileal mucosal barrier dysfunction induced by subjecting rats to profound HS for 2 hours. The most active compound in this assay, XJB-5-131, ameliorated peroxidation of the mitochondrial phospholipid, cardiolipin, in ileal mucosal samples from rats subjected to HS. XJB-5-131 also ameliorated HS-induced activation of the pro-apoptotic enzymes, caspases 3 and 7, in ileal mucosa. Intravenous treatment with XJB-5-131 (2 micromol/kg) significantly prolonged the survival of rats subjected to profound blood loss (33.5 mL/kg) despite administration of only a minimal volume of crystalloid solution (2.8 mL/kg) and the absence of blood transfusion. These data support the view that mitochondrially targeted electron acceptors and SOD mimics are potentially valuable therapeutics for the treatment of serious acute conditions, such as HS, which are associated with marked tissue ischemia.

Supplementation of dietary enzymes has varying effects on apparent protein and amino acid digestibility in reproducing sows

The effects of two enzyme (Enz) products on ileal and total tract digestibility (ID and TTD) were evaluated in sows ( n = 8) fed corn–soybean meal diets. Treatments were: 1) control diet; 2) control + Enz 1 (protease and cellulase activity); and 3) control + Enz 2 (xylanase activity). Ileal and fecal samples were collected at week 6–7 and 12–13 of gestation and week 2–3 of lactation. There were no improvements in digestibility during gestation. The ID% of dry matter (DM: 77.3, 79.3, 81.7) and N (81.2, 82.5, 84.3) as well as TTD% of DM (89.8, 90.7, 90.8) and N (89.0, 90.0, 90.6) during lactation were affected to varying degrees by the enzymes. The TTD of DM ( P = 0.05) and N ( P = 0.10) were positively affected by Enz 1. The ID of DM ( P = 0.01) and N ( P = 0.02) as well as TTD of DM ( P = 0.05) and N ( P = 0.02) were improved by Enz 2. The improvement in ID of N was not associated with any particular amino acids (AA) but represented small increases in ID of 8 of 10 essential AA with Enz 1 ( P = 0.04) and 10 of 10 for Enz 2 ( P = 0.01). The product containing xylanase activity, in particular, appears to have potential to increase digestibility of nutrients during lactation in sows.

Propolis reduces bacterial translocation and intestinal villus atrophy in experimental obstructive jaundice

To investigate the effects of propolis on bacterial translocation and ultrastructure of intestinal morphology in experimental obstructive jaundice. Thirty Wistar-Albino male rats were randomly divided into three groups, each including 10 animals: group I, sham-operated; group II, ligation and division of the common bile duct (BDL); group III, BDL followed by oral supplementation of propolis 100 mg/kg per day. Liver, blood, spleen, mesenteric lymph nodes, and ileal samples were taken for microbiological, light and transmission electron microscopic examination on postoperative 7th d after sacrification. The mean number of villi per centimeter and mean mucosal height of the propolis group were significantly different in the BDL group (P = 0.001 and 0.012, respectively). The electron microscopic changes were also different between these groups. Sham and BDL + propolis groups had similar incidence of bacterial translocation (BT). The BDL group had significantly higher rates of BT as compared with sham and BDL + propolis groups. BT was predominantly detected in MLNs and the most commonly isolated bacteria was Escherichia coli. Propolis showed a significant protective effect on ileal mucosa and reduced bacterial translocation in the experimental obstructive jaundice model. Further studies should be carried out to explain the mechanisms of these effects.

A Comparison of Methods to Determine Amino Acid Digestibility of Feed Ingredients for Chickens

Two experiments were conducted to compare standardized ileal amino acid (AA) digestibility in 7- and 21-d-old chicks and true AA digestibility as determined by the precision-fed cecectomized rooster assay for several ingredients used in poultry feeds. Diets were formulated to contain soybean meal, cottonseed meal, poultry by-product meal, and fish meal in experiment 1 and corn, wheat, soybean meal, poultry by-product meal, feather meal, and fish meal in experiment 2 as the sole sources of protein. Celite, used as an indigestible marker, was added at 1.5% of the diet. The test diets were fed ad libitum to broiler chicks from 0 to 7 d of age in experiment 1 and from 4 to 7 and 17 to 21 d of age in experiment 2. Ileal digesta samples were collected after euthanizing the birds at 7 d of age in experiment 1 and at 7 and 21 d of age in experiment 2. Additionally, cecectomized, Single-Comb White Leghorn roosters were used for crop intubation of the test diets for determination of true AA digestibility. In experiment 1, AA digestibility of all ingredients tested was significantly lower at 7 d of age than when determined by the rooster assay. In experiment 2, no differences were detected between AA digestibility at 7 or 21 d of age in the chick assay for the majority of the indispensable AA. However, the AA digestibility coefficients obtained by the chick assay at 7 d and, in some cases, at 21 d of age, were significantly lower than those obtained by the rooster assay. In conclusion, there were differences in the AA digestibility coefficients obtained through the chick and the rooster assays. Such discrepancies could be associated with an age effect or the methodological differences between both methods.

Effects of fructan supplementation on intestinal microbial populations of salmonella‐challenged weanling puppies

Weanling puppies are at high risk for illness if exposed to Salmonella. Our objective was to evaluate the effects of fructan consumption on changes in fecal, colonic digesta, and ileal mucosa microbiota populations in weanling puppies challenged with Salmonella. After a 1 wk baseline period, 30 hound-cross puppies (12 wk-old) were randomly assigned to three diets containing: no prebiotic (control); 1% short-chain fructooligosaccharides (scFOS); or 1% inulin. After 14 d on dietary treatment, dogs received an oral gavage of: S. typhimurium DT104 (5×108 CFU); or 0.9% saline (n = 5 per treatment). Fresh feces were collected at baseline and d 14. Colonic digesta and ileal mucosa samples were collected 48 h post-infection. Following DNA extraction, denaturing gradient gel electrophoresis (DGGE) was used to generate microbial fingerprints and qPCR was used to quantify Bifidobacterium, Lactobacillus, C. perfringens, E. coli, and Salmonella concentrations. Microbial diversity was determined by calculating Dice’s coefficient. Data were analyzed using the mixed method of SAS. Although E. coli and C. perfringens were unchanged due to treatment, changes in Lactobacillus concentrations were greater (P < 0.05) in puppies fed inulin. It not known if this change contributed to the apparent protection from ileal mucosal damage that was observed in puppies fed inulin. Funding: USDA-CSREES (Project #ILLU-538-396).

True phosphorus digestibility and the endogenous phosphorus outputs associated with brown rice for weanling pigs measured by the simple linear regression analysis technique

The objectives of this study were to determine true phosphorus (P) digestibility, degradability of phytate-P complex and the endogenous P outputs associated with brown rice feeding in weanling pigs by using the simple linear regression analysis technique. Six barrows with an average initial body weight of 12.5 kg were fitted with a T-cannula and fed six diets according to a 6 × 6 Latin-square design. Six maize starch-based diets, containing six levels of P at 0.80, 1.36, 1.93, 2.49, 3.04, and 3.61 g/kg per kg dry-matter (DM) intake (DMI), were formulated with brown rice. Each experimental period lasted 10 days. After a 7-day adaptation, all faecal samples were collected on days 8 and 9. Ileal digesta samples were collected for a total of 24 h on day 10. The apparent ileal and faecal P digestibility values of brown rice were affected ( P < 0.01) by the P contents in the assay diets. The apparent ileal and faecal P digestibility values increased from − 48.0 to 36.7% and from − 35.6 to 40.0%, respectively, as P content increased from 0.80 to 3.61 g/kg DMI. Linear relationships ( P < 0.05), expressed as g/kg DMI, between the apparent ileal and faecal digestible P and dietary levels of P, suggested that true P digestibility and the endogenous P outputs associated with brown rice feeding could be determined by using the simple regression analysis technique. There were no differences ( P>0.05) in true P digestibility values (57.7 ± 5.4 v. 58.2 ± 5.9%), phytate P degradability (76.4 ± 6.7 v. 79.0 ± 4.4%) and the endogenous P outputs (0.812 ± 0..096 v. 0.725 ± 0.083 g/kg DMI) between the ileal and the faecal levels. The endogenous faecal P output represented 14 and 25% of the National Research Council (1998) recommended daily total and available P requirements in the weanling pig, respectively. About 58% of the total P in brown rice could be digested and absorbed by the weanling pig. Our results suggest that the large intestine of the weanling pigs does not play a significant role in the digestion of P in brown rice. Diet formulation on the basis of total or apparent P digestibility with brown rice may lead to P overfeeding and excessive P excretion in pigs.

Effects of Feed Additives and Mixed Eimeria Species Infection on Intestinal Microbial Ecology of Broilers

Evaluation of digestive microbial ecology is necessary to understand effects of growth-promoting feed. In the current study, the dynamics of intestinal microbial communities (MC) were examined in broilers fed diets supplemented with a combination of antibiotic (bacitracin methylene disalicylate) and ionophore (Coban 60), and diets containing 1 of 2 essential oil (EO) blends, Crina Poultry (CP) and Crina Alternate (CA). Five treatments were analyzed: 1) unmedicated uninfected control; 2) unmedicated infected control; 3) feed additives monensin (bacitracin methylene disalicylate) + monensin (Coban 60; AI); 4) EO blend CP; and 5) EO blend CA. Additives were mixed into a basal feed mixture, and EO were adjusted to 100 ppm. Chicks were infected by oral gavage at 19 d of age with Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Duodenal, ileal, and cecal samples were taken from 12 birds per treatment just before and 7 d after challenge; 2 samples each were pooled to give a final number of 6 samples total; and all pooled samples were frozen until used for DNA extraction. Denaturing gradient gel electrophoresis was used to examine PCR-amplified fragments of the bacterial 16S ribosomal DNA variable region. Results are presented as percentages of similarity coefficients (SC). Dendrograms of PCR amplicon or band patterns indicated MC differences due to intestinal location, feed additives, and cocci challenge. Essential oil blends CP and CA affected MC in all gut sections. Each EO had different effects over MC, and they differed in most instances from the AI group. The cocci challenge caused drastic MC population shifts in duodenal, ileal, and cecal sections (36.7, 55.4, and 36.2% SC, respectively). Diets supplemented with CP supported higher SC between pre- and postchallenge MC (89.9, 83.3, and 76.4%) than AI (81.8., 57.4, and 60.0%). We concluded that mixed coccidia challenge caused drastic shifts in MC. These EO blends modulated MC better than AI, avoiding drastic shifts after a mixed challenge.

Influence of the amount of dietary fiber on the available energy from hindgut fermentation in growing pigs: Use of cannulated pigs and in vitro fermentation1

Available energy from hindgut fermentation to pigs fed various amounts of dietary fiber was investigated using an in vivo-in vitro methodology. Six growing pigs fitted with a simple T-shaped cannula at the terminal ileum, and following a Latin-square design, were fed 3 diets differing in the content of non-starch polysaccharides (NSP): a low fiber diet (LFD, 77 g/kg of DM), a standard fiber diet (SFD, 160 g/kg of DM), and a high fiber diet (HFD, 240 g/kg of DM). After adaptation to the diet for 10 d, samples from feces and ileum were collected and analyzed for DM, energy, NSP, and chromic oxide; feces were also analyzed for short chain fatty acids (SCFA). Freeze-dried ileal samples (10 g/L) were fermented in vitro in a fecal slurry consisting of an anaerobic mineral salt medium and feces (50 g/L) from cannulated pigs fed the same diets. Available energy was calculated from the amount of SCFA produced in vitro after 48 h of incubation. Nonstarch polysaccharide content in the fermented material was measured to assess the in vitro degradation of this fraction. Increasing dietary NSP from 77 to 240 g/kg of feed DM increased (P < 0.001) ileal flow from 199 to 468 g/kg of feed, leading to a reduction in the energy digested at the terminal ileum, from 15 to 11 MJ/kg of feed DM and an increment in energy digested in the hindgut, from 1.6 to 3.5 MJ/kg of feed DM. Total in vitro production of SCFA/kg of feed DM was dependent on the amount of ileal substrate available for fermentation; that is, increased concentrations of NSP in the diet led to an increase in the SCFA that may be available to the animal (P < 0.001). The molar ratio of SCFA produced in vitro was affected by diet; the high fiber diet showed the greatest (P = 0.004) proportion of acetic acid, and the low fiber diet showed a tendency (P = 0.081) to an increased butyric acid proportion compared with the other 2 diets. Net disappearance of NSP during fermentation in vivo and in vitro were compared and showed a close relationship (P < 0.001, slope = 0.906, r = 0.960). In our experimental conditions, available energy as SCFA to the animal from hindgut fermentation increased with the concentration of dietary NSP (P < 0.001) and provided between 7.1 and 17.6% of the total available energy.

Effects of Prenatal Lipopolysaccharide Exposure on Epithelial Development and Function in Newborn Rat Intestine

Maternal infection during pregnancy is associated with several neonatal morbidities, including periventricular leukomalacia and lung maldevelopment and injury. To test the hypothesis that responses to prenatal maternal exposure to lipopolysaccharide (LPS) alter intestinal epithelial development and function in newborn rats. Timed-pregnancy female Sprague-Dawley rats were administered either 2 mg LPS or an equal volume of isotonic saline by intraperitoneal injection at E16 and allowed to deliver naturally. Pups were weighed and then killed at days of life (DOL) 0, 3, 7 and 14. Morphometric parameters were measured on standard hematoxylin and eosin-stained sections using ImagePro software. Immunohistochemistry was performed with antibody specific for inducible nitric oxide synthase (iNOS) and 3-nitrotyrosine on distal ileal intestinal samples analyzed at each time point. Optical density was determined and quantified for site-specific regions of intestinal sections. On DOL 14, in vivo mucosal permeability was measured by feeding rats fluorescein isothiocyanate (FITC) via orogastric tube; and then serum FITC was measured. There were no significant differences in pup weights. Mucosal thicknesses were significantly less in the distal ileum from pups born to LPS-exposed dams on DOL 0, 3 and 7 (P < 0.001). On DOL 0, iNOS protein concentrations in the prenatal LPS treatment group were significantly greater than iNOS protein concentrations in the distal villus (P < 0.001), proximal villus/crypts (P < 0.01), submucosa (P < 0.001) and muscularis (P < 0.01) in the distal small intestine of the control group. On DOL 3, 7 and 14, significant differences were observed in iNOS protein concentrations in the distal villus and submucosal regions between groups (P < 0.001). On DOL 0, 3, 7 and 14, 3-nitrotyrosine immunostaining was significantly elevated in the prenatal LPS-exposed pups in the distal villus on (P < 0.001) as well as in the submucosa on DOL 3 (P < 0.001). Serum FITC measurement was significantly greater in prenatal LPS exposure group at DOL 14 (P < 0.001). Maternal exposure to LPS during pregnancy alters intestinal growth and regulation of iNOS in the newborn rat intestine.

Effect of dietary inclusion of distillers dried grains with solubles, soybean hulls, or a polyclonal antibody product on the ability of growing pigs to resist a Lawsonia intracellularis challenge1

An experiment was conducted to determine if dietary inclusion of distillers dried grains with solubles (DDGS), soybean hulls, or soybean hulls sprayed with an egg-based, polyclonal antibody product would reduce the incidence or severity of infection, or both, in growing pigs after a Lawsonia intracellularis challenge. One hundred 17-d-old weaned pigs were blocked by sex, ancestry, and BW, and randomly allotted to 1 of 5 treatment groups: negative control, unchallenged, corn-soy diet; positive control, challenged, corn-soy diet; 20% DDGS diet (D), challenged; 5% soybean hulls diet (SH), challenged; and SH sprayed with a polyclonal antibody product diet, challenged. Challenged pigs were orally inoculated with 6.4 x 10(8) L. intracellularis organisms after a 4-wk prechallenge feeding period. On d 21 postchallenge, pigs were euthanized, lesions of intestinal mucosa were evaluated, and ileal tissue samples were analyzed by immunohistochemistry to determine the presence and proliferation rate of L. intracellularis. Challenging pigs with L. intracellularis reduced growth rate, feed intake, and efficiency of gain (P < 0.02) and increased the proportion of internal organ weights relative to BW (P < 0.01). Dietary treatment did not affect growth performance pre- or postchallenge (P > 0.10). Heart, empty stomach, and liver weights were similar among dietary treatments (P > 0.10). Weight of the large intestine as a percentage of BW was increased in D and SH pigs compared with positive control pigs (P < 0.05). Lesion length, prevalence, and severity, and fecal shedding of L. intracellularis were primarily unaffected by dietary treatment (P > 0.10), although ileal lesion length and severity observed tended to be greater in the SH sprayed with polyclonal antibody product diet vs. the D pigs (P < 0.10). Results from a previous study indicated that diet composition may affect length, severity, and prevalence of lesions caused by L. intracellularis in growing pigs subjected to a moderate challenge. However, beneficial results were not observed by feeding the dietary treatments used in this study.

Effect of dietary inclusion of distillers dried grains with solubles on the ability of growing pigs to resist a Lawsonia intracellularis challenge1

An experiment was conducted to determine if including distillers dried grains with solubles (DDGS) in the diet of growing pigs reduces the incidence or severity of infection after a Lawsonia intracellularis challenge. Eighty 17-d-old weaned pigs were blocked by sex, ancestry, and BW and randomly allotted to 1 of 4 treatment groups: negative control (NC), unchallenged, corn-soy diet; positive control (PC), challenged, corn-soy diet; 10% DDGS diet (10D), challenged; and 20% DDGS diet (20D), challenged. Challenged pigs were orally inoculated with 1.5 x 10(9) L. intracellularis organisms after a 4-wk prechallenge feeding period. On d 21 postchallenge, pigs were euthanized, lesions of intestinal mucosa were evaluated, and ileal tissue samples were analyzed by immunohistochemistry to determine the presence and proliferation rate of L. intracellularis. Compared with unchallenged pigs, challenging pigs with L. intracellularis reduced growth rate, feed intake, and efficiency of gain (P < 0.01) and increased gauntness (P < 0.05) and diarrhea (P < 0.01). Diet did not affect growth performance postchallenge (P > 0.40). Feeding 10 or 20% DDGS diets did not reduce lesion length, prevalence, proliferation of L. intracellularis, or severity of lesions (P > 0.10). Thus, dietary inclusion of DDGS did not reduce the incidence or severity of lesions under the conditions of a severe L. intracellularis challenge used in this study.

Effect of including distillers dried grains with solubles in the diet, with or without antimicrobial regimen, on the ability of growing pigs to resist a Lawsonia intracellularis challenge1

A disease challenge experiment was conducted to determine if including 10% dried distillers grains with solubles (DDGS) in the diet, with or without antimicrobial supplementation, reduces the incidence or severity, or both, of intestinal lesions in growing pigs after an Lawsonia intracellularis challenge. One hundred 17-d-old weaned pigs were blocked by sex, ancestry, and BW and randomly allotted to 1 of 5 treatment groups: negative control, unchallenged, corn-soy diet; positive control, challenged, corn-soy diet; 10% DDGS diet, challenged; positive control with antimicrobial regimen, challenged; and 10% DDGS diet with antimicrobial regimen, challenged. For antimicrobial-supplemented treatments, diets contained 33 ppm bacitracin methylene disalicylate throughout the experiment, with chlortetracycline (Aureomycin) pulsed at 550 ppm from d 3 prechallenge to d 11 postchallenge. Challenged pigs were orally inoculated with 8.0 x 10(8) L. intracellularis organisms after a 4-wk prechallenge period. On d 21 postchallenge, pigs were euthanized, lesions of intestinal mucosa were evaluated, and ileal tissue samples were analyzed by immunohistochemistry to determine the presence and proliferation rate of L. intracellularis. Compared with other dietary treatments, feeding a diet containing 10% DDGS reduced ileum and colon lesion length and prevalence (P < 0.05) and reduced severity of lesions in the ileum (P < 0.05) and colon (P < 0.10) in challenged pigs. Compared with other challenged pigs, those fed the diet containing the antimicrobial regimen had a lower prevalence and severity of lesions in the jejunum (P < 0.05) and tended to have reduced total tract lesion length (P = 0.11). Compared with other challenged pigs, pigs on the 10% DDGS diet with antimicrobial regimen exhibited no differences in length, severity, or prevalence of lesions (P > 0.15), but fecal shedding of L. intracellularis was reduced on d 14 postchallenge (P < 0.05). No dietary effects on fecal shedding were observed by d 20 postchallenge (P > 0.10). The proportion of cells infected with L. intracellularis was reduced when DDGS (P = 0.05) or antimicrobial (P = 0.10) diets were fed. Under the conditions of this experiment, dietary inclusion of 10% DDGS appears to provide some benefit to growing pigs subjected to a moderate L. intracellularis challenge, similar to those of a currently approved antimicrobial regimen.

Mycotoxin fumonisin B 1 selectively down-regulates the basal IL-8 expression in pig intestine: in vivo and in vitro studies

Fumonisin B 1 (FB 1) is a mycotoxin produced by Fusarium verticillioides and F. proliferatum, common contaminants of maize. FB 1 causes toxicological effects in laboratory and domestic animals including pigs. The gastrointestinal tract represents a barrier encountered by exogenous food compounds. The purpose of our study was to determine FB 1 effects on intestinal immune response. Nine recently weaned piglets orally received 0.5 mg of purified toxin/kg of body weight/day for 7 days, while eight other animals were kept as controls. After necropsy, ileal samples were analyzed for five pro-inflammatory cytokines mRNA expression by RT-PCR. No difference was observed for IL-1β, IL-6, IL-12 and TNF-β mRNA levels between control and FB 1-treated animals. In contrast, FB 1 treatment induced a significant down-regulation of the expression of IL-8 mRNA in the pig ilea. The effect of FB 1 on the IL-8 expression was also examined in the porcine intestinal epithelial cell line IPEC-1. FB 1 decreases the expression of IL-8, both at the mRNA and protein levels, in a dose-dependant manner. Taken together, our data demonstrate that FB 1 alters the intestinal immune response by decreasing the level of IL-8. This may contribute to the increased intestinal colonization by pathogenic Escherichia coli that was observed in FB 1-treated pigs and may have implications for humans/animals consuming FB 1-contaminated food/feed.

Interstitial cells of Cajal and electrical activity of smooth muscle in porcine ileum

To identify the interstitial cells of Cajal in the porcine ileum for the first time immunohistochemically and to examine the electrical properties of intestinal smooth muscle in the same region. In vitro intracellular microelectrode recordings were made from smooth muscle cells in cross-sectional preparations from abattoir-derived healthy porcine ileum. Immunohistochemical labelling of interstitial cells of Cajal was performed using an anti-Kit antibody. Slow waves were recorded in the circular muscle layer of all ileal preparations. The mean resting membrane potential of smooth muscle cells was -61.0 +/- 1.3 mV. Slow waves had a mean amplitude of 8.5 +/- 0.5 mV, a frequency of 9.9 +/- 0.1 cycles per minute and a duration of 5.6 +/- 0.1 s. A waxing and waning pattern of slow wave activity was occasionally observed. In addition, higher frequency spiking activity associated with contractions was observed in some recordings. The L-type calcium channel blocker nifedipine abolished both the spiking activity and the contractions, but had no significant effect on slow wave characteristics. Current-injection manipulation of the resting membrane potential had no effect on slow wave amplitude, frequency or duration. Kit-immunoreactive interstitial cells of Cajal were identified in the ileal samples and were present in the region of the myenteric plexus and in the circular and longitudinal muscle layers. This study recorded slow waves in vitro and demonstrated immunohistochemically the presence of interstitial cells of Cajal in the normal porcine ileum. This study forms a basis for future physiological and pathophysiological comparative studies of intestinal motility.

The Effect of Source of Dietary Fiber and Starch on Ileal and Fecal Amino Acid Digestibility in Growing Pigs

Studies were carried out with a repeated 4x4 Latin square design with eight cannulated pigs fed four experimental diets to investigate the effect of dietary fiber and starch sources on apparent ileal and fecal amino acid digestibility. Each period lasted 15 d, with diet acclimation from d 1 to 7, feces collection for 48 h on d 8 to 9 and ileal sample collection for 12 h on d 13 to 15. The four experimental diets consisted mainly of cooked rice with the addition of protein sources (CON), partial replacement of cooked rice with either potato starch (PS), sugar beet pulp (SBP) or wheat bran (WB). Chromic oxide was used as an indigestible marker. With the exception of histidine, lysine and tryptophan, no differences were observed in the apparent ileal digestibility of amino acids between diets. The inclusion of potato starch did not affect the ileal and fecal amino acid digestibility. In comparison with diet CON, a decreased (p<0.05) ileal digestibility of histidine was found in pigs fed diet SBP, while the ileal digestibilities of histidine, lysine and tryptophan were decreased (p<0.05) by the inclusion of wheat bran. Inclusion of fiber sources (sugar beet pulp and wheat bran) caused a reduction (p<0.05) in the fecal amino acid digestibility and the net disappearance of amino acids in the large intestine. Of the indispensable amino acids, there was a 'net synthesis' for methionine in the large intestine of pigs when diets were supplemented with dietary fiber. The decrease in fecal amino acid digestibility with the addition of dietary fiber indicates an increase in the synthesis of bacterial protein in the large intestine.

Intestinal Microbial Ecology of Broilers Vaccinated and Challenged With Mixed Eimeria Species, and Supplemented with Essential Oil Blends

Intestinal microbiota is an important component in the development of defense mechanisms in the gut mucosa. This project determined the dynamics of intestinal microbial communities (MC) of broilers vaccinated at first day of age with live oocysts of Eimeria species and fed diets supplemented with 2 specific essential oil (EO) blends, Crina Poultry (CP) and Crina Alternate (CA). Five treatments were analyzed: 1) unmedicated-uninfected (UU) control; 2) unmedicated-infected (UI) control; 3) vaccinated with Advent cocci-vaccine and without feed additive (COV) supplements; 4) vaccinated with Advent and supplemented with CP; and 5) vaccinated with Advent and supplemented with CA. The EO blends were added at 100 ppm to the same basal diets. Chicks were gavage-infected at 19 d of age with Eimeria acervulina, Eimeria maxima, and Eimeria tenella. Duodenal, ileal, and cecal samples were taken from 12 birds per treatment just before the infection and 7 d after the challenge, pooled in 6 samples, and frozen. Denaturing gradient gel electrophoresis was used to examine PCR-amplified fragments of the bacterial 16S ribosomal DNA variable region. Results are presented as percentages of similarity coefficients (SC). Dendrograms of amplicon patterns indicated MC differences due to intestinal location, feed additives, and cocci infection. The EO blends CP and CA did affect MC in all gut sections. The cocci-infection caused drastic MC population shifts in duodenal, ileal, and cecal sections (36.7, 55.4, and 36.2% SC, respectively). The CP-supplemented birds had higher SC between pre- and postchallenge MC in duodenal and ileal (73.3, 81.8%) than COV (66.4, 66.5%). However, COV broilers had the smallest changes in cecal MC after infection (79.5% SC). We concluded that cocci-vaccination causes small changes in intestinal MC, but challenge causes drastic shifts. The EO blend supplementation modulates MC in cocci-vaccinated broilers, avoiding drastic shifts after a mixed coccidia infection. Correlations between MC dynamics and host responses are discussed.

Effect of feeding schedule on apparent energy and amino acid digestibility by growing pigs

An experiment was conducted to investigate the effects of feeding schedule on dry matter (DM), gross energy (GE), crude protein (CP), and amino acid (AA) digestibility by growing pigs. Six growing castrates (initial body weight: 40.5 ± 2.25 kg) were equipped with a T-cannula in the distal ileum and allotted to a repeated 3 × 3 Latin square design with three animals and three periods in each square. A maize–soybean meal based diet (181 g CP/kg) was formulated and fed to the pigs in three different feeding schedules; i.e., one meal per day, two meals per day, or free access to feed. Pigs fed one or two meals per day were allotted a daily amount of feed equivalent to three times the maintenance energy requirement. Fecal and ileal samples were collected from all pigs in each period. The coefficients of ileal apparent digestibility (CIAD) for DM, GE, CP, and AA were calculated as were the coefficients of total tract apparent digestibility (CTTAD) of DM and GE. Results of the experiment showed that there were no differences in the CIAD for DM, GE, or for any of the AA among the three feeding schedules. In contrast, pigs given free access to the diet had lower (P<0.002) CTTAD for DM and GE compared with pigs fed a restricted amount of feed either once or twice daily. Therefore, the concentration of digestible energy in the diet was lower if calculated from pigs given free access to the diet compared with pigs fed the diet once or twice daily (14.38 MJ/kg versus 14.83 MJ/kg and 15.13 MJ/kg). It is concluded that the feeding schedule does not influence the CIAD for DM, GE, CP, or AA by pigs. Therefore, any of the three feeding schedules that were used in the current experiment may be used in experiments aimed at measuring CIAD in pigs. However, if the CTTAD of DM and GE are measured, it is recommended that pigs are given free access to feed.

The digestive fate of Escherichia coli glutamate dehydrogenase deoxyribonucleic acid from transgenic corn in diets fed to weanling pigs1

Corn containing genetically engineered plasmid DNA encoding an Escherichia coli glutamate dehydrogenase (gdhA) was fed to 19-d-old weanling swine to trace the digestive fate of the transgenic DNA. Eight pens of 8 pigs were fed a commercial (nongdhA) starter for 2 wk. One pig was randomly selected from each pen for 0-h control samples. The remaining 56 pigs were transitioned onto a corn-soybean meal diet and fed a diet containing 58% gdhA corn for approximately 1 wk; immediately thereafter, liver, 10th rib muscle, white blood cells, and plasma from the hepatic portal vein and ingesta from the stomach, distal ileum, and large intestine were collected. The DNA was extracted and the concentration determined via spectrophotometry. Polymerase chain reaction and gel electrophoresis were performed with primers designed to amplify 490 bp that included the plasmid's ligation site between the maize ubiquitin and the gdhA genes. The gdhA corn-derived DNA and diet served as positive assay controls, and conventional corn DNA and distilled water acted as negative assay controls. Detection limits were 0.99 fg of target DNA confounded with 500 ng of conventional corn DNA per each 20 &L reaction. Transgenic DNA was detected in 71.43% of the stomach and 1.79% of the ileal ingesta samples from treatment animals but was not detected in the large intestine, white blood cells, plasma, liver, or muscle samples. Transgenic DNA was not detected in any sample from 0-h control animals. Stomach and ileal ingesta samples were further analyzed using real-time PCR. With an estimated limit of detection of 1.049 ag/microL, 89.29% of the stomach ingesta samples were positive (average 1.56 fg target DNA). The proportion of transgenic DNA to total DNA differed between diet and stomach ingesta samples (P < 0.001). Despite the greater sensitivity of real-time PCR, target DNA was detected in only 1.79% of ileal ingesta. These data suggest that the gdhA transgene began degradation in the stomach and was nondetectable in the large intestine.

Post‐natal development of the porcine microbiota composition and activities

The current study describes the development of the porcine microbiota and its metabolic activities during the neonatal and weaning period. Using 16S rRNA-based approaches, we first analysed the ileal and colonic microbiota of neonatal piglets at days 2, 5 and 12 after birth. To further investigate the effect of weaning at 3 weeks of age, 19-day-old piglets (n = 64) were randomly allocated into two groups. Half of the piglets remained with their sows throughout the study, while the remaining piglets were weaned. As revealed by sequence analysis of 16S rRNA gene amplicons, the samples of 2-day-old piglets harboured a consortium of bacteria related to Escherichia coli, Shigella flexneri, Lactobacillus sobrius, Lactobacillus reuteri and Lactobacillus acidophilus. Moreover, species-specific real-time polymerase chain reaction assays unveiled that L. sobrius and L. reuteri predominated in the ileal samples of the neonatal and unweaned piglets with population levels up to 7 x 10(8) cells per gram of lumen content. Following weaning, however, these two lactobacilli were detected at significantly lower levels (< 10(3)) in the ileal samples. Furthermore, a shift in composition and metabolic activities of the predominant microbiota, and emergence of clostridia and E. coli, were encountered in the intestinal samples of the piglets after the early post-weaning period.