Molecular characterisation of fructose transport in equine small intestine

REASONS FOR PERFORMING STUDY: Fructose can be a suitable carbohydrate supplement for horses before and/or during endurance exercise. In comparison to glucose, the ingestion of fructose results in a lower insulin peak and less marked fluctuations in blood glucose during exercise, potentially avoiding hypoglycaemia-induced exhaustion. OBJECTIVES: To assess the capacity of the equine small intestine to absorb fructose and to determine the mechanism, molecular structure and properties of equine intestinal fructose transport. METHODS: Using PCR-based strategies, RNA isolated from equine small intestine and primers designed to homologous regions of the fructose transporter, GLUT5, cDNA of other species, we cloned and sequenced equine GLUT5 (eGLUT5). Northern and western blot analyses, in conjunction with immunohistochemistry, utilising eGLUT5 cDNA and antibodies, assessed expression of eGLUT5 along the longitudinal and radial axes of the small intestine. Functional properties of fructose transport in intestinal brush-border membrane vesicles were measured using the rapid-filtration technique. RESULTS: eGLUT5 is expressed in the villus enterocytes with highest levels in duodenum>jejunum and lowest in the ileum. Kinetic studies indicate eGLUT5 is a low affinity, high capacity transporter. CONCLUSIONS: Equine small intestine has the capacity to absorb fructose. POTENTIAL RELEVANCE: The molecular probes produced in these studies can be used as diagnostic aids to determine equine intestinal monosaccharide malabsorption.     

Gentamicin tissue concentrations in equine small intestine and large colon

Gentamicin sulfate (2.2 mg/kg of body weight, IV) was given to anesthetized horses. Jejunal and large colon tissue samples (1 g), serum, and urine were collected over a 4-hour period. Maximum gentamicin concentrations in serum (10.06 +/- 2.85 micrograms/ml) occurred at 0.25 hours after injection. Maximum gentamicin concentrations in the large colon (4.13 +/- 1.80 micrograms/ml) and jejunum (2.26 +/- 1.35 micrograms/ml) occurred in horses at 0.5 and 0.33 hours, respectively. Tissue concentrations decreased in parallel with serum concentrations and were still detectable at the end of the 4-hour period. During the time that samples were collected, the total amount of gentamicin excreted in the urine ranged from 7.21 +/- 3.11 mg to 11.91 +/- 7.12 mg, with a mean urinary concentration of 57.01 +/- 5.37 micrograms/ml. Over the 4-hour collection period, the fraction of dose that was excreted unchanged in the urine was 4.8 +/- 1.9%. Pharmacokinetic analyses of the serum concentration-time data gave a serum half-life of 2.52 +/- 1.29 hours, volume of distribution of 227 +/- 83 ml/kg, and body clearance of 1.12 +/- 0.26 ml/min/kg. The half-lives of the antibiotic in the jejunum and large colon were 1.32 and 1.33 hours, respectively.     

Limits to starch digestion in the ruminant small intestine

Site and extent of starch digestion by ruminant animals varies with species, grain type and processing method. Based on a review of 40 different experiments with cattle, between 18 and 42% of the dietary starch from corn and sorghum grains fed to cattle reaches the small intestine for digestion. With more extensive grain processing, a smaller quantity of starch reaches the small intestine. In the small intestine, from 47 to 88% of the presented starch is digested, while in the large intestine, 33 to 62% of the presented starch is digested. Though limits to digestion in and absorption from the small intestine can be demonstrated by infusing starch and glucose into the duodenum, enzymatic capacity does not appear to limit intestinal starch digestion since no plateau in the amount of starch disappearing from the small intestine is detected with typical diets. Yet, extent of digestion is incomplete. Other factors, such as time and surface exposure may limit small intestinal digestion of starch. Processing methods to reduce particle size or alter the protein matrix, which cements starch granules together, will increase the extent of digestion both in the rumen and in the small intestine. Performance data from growing cattle fed processed corn and sorghum grains indicate that starch was used 42% more efficiently if it was digested in the small intestine rather than in the rumen. Though total tract starch digestibility is of primary concern, results support the concept that energetic efficiency of growing ruminants is greater if starch is digested in the small intestine rather than in the rumen.     

单胃动物小肠内可消化性碳水化合物的吸收及其调控

本文阐述单胃动物小肠内可消化性碳水化合物的吸收及其调控机制。可消化性碳水化合物在单胃动物体内的降解产物主要为葡萄糖、果糖、半乳糖和双糖。葡萄糖、果糖和半乳糖在小肠内,由小肠内绒毛上皮细胞或细胞间隙直接吸收;双糖在双糖酶的作用下水解成单糖形式,为小肠绒毛上皮细胞吸收。其吸收机制可分为3个途径:主动吸收、被动吸收以及通过细胞间隙直接吸收。其中主动吸收是主要的吸收途径。调控小肠可消化性碳水化合物吸收的因素较多,包括吸收面积、Na 电化学梯度、细胞膜脂质成分、转运细胞与非转运细胞比例、转运子周转速率、亲和系数等因素。通过多种因子的调控作用,能有效地促进碳水化合物的吸收,以满足动物体的生长和发育的需要。     

Eimeria organisms develop in the epithelial cells of equine small intestine

Histopathologic and immunohistochemical examinations were performed to determine the origin of host cells parasitized by Eimeria in the small intestines collected from five foals. Eimeria organisms at various stages (mainly microgametes and macrogametes) were frequently found in the cytoplasm of hypertrophied host cells in the lamina propria at the tips of villi of the jejunum and ileum. The cytoplasm of the host cell was immunohistochemically positive for cytokeratin AE1/AE3 and cytokeratin 13 and was negative for vimentin, desmin, alpha-smooth muscle actin, chromogranin A, neuron-specific enolase, and factor VIII. The host cells parasitized by Eimeria species had the immunostaining characteristics of epithelial cells but not of mesenchymal cells, endothelial cells of lacteals or capillaries, smooth muscle cells or neuroendocrine cells. These results suggest that the host cell of Eimeria species is possibly derived from intestinal epithelial cells and then displaced into the lamina propria of the small intestine.     

Post operative neutrophilic inflammation in equine small intestine after manipulation and ischaemia

REASONS FOR PERFORMING STUDY: Post operative ileus (POI) remains an important cause of post operative morbidity and mortality in the horse. However, clinical progression of naturally occurring cases of POI in both horse and man does not entirely support the 'neurogenic' hypothesis as the sole mechanism of POI; and the hypothesis that inflammation plays a major role at 12-24 h after surgery requires validation. HYPOTHESIS: An inflammatory infiltrate in the muscularis externa and myenteric plexus of equine jejunum is present 18 h following a period of ischaemia. METHODS: Samples of normal jejunum, jejunum from the proximal resection margins of clinical cases and jejunum obtained 18 h after 1 or 2 h ischaemia or manipulation alone were evaluated for neutrophil infiltration. Samples obtained 18 h after surgery were additionally evaluated for leucocyte activation using calprotectin immunohistochemistry. Results were evaluated by ANOVA and P < 0.05 was considered significant. RESULTS: Significant neutrophilic inflammation was identified in the samples from the proximal resection margins of clinical cases compared to uninjured jejunum. In experimental cases, neutrophilic inflammation appeared to be increased further by 18 h and was identified through all intestinal layers, particularly in the serosa, fascial planes around circular and longitudinal muscle fibres, and myenteric plexus. This elevated level of neutrophilic inflammation was mirrored by an increased number of calprotectin-positive cells in these intestinal layers, indicating leucocyte activation. CONCLUSIONS: Significant neutrophilic inflammation occurs in equine jejunal myenteric layers 18 h after surgery. POTENTIAL RELEVANCE: This neutrophilic inflammation coincides with the clinical time point at which POI is identified and may indicate that inflammatory pathways, rather than solely neurogenic pathways, are responsible for POI in the horse.     

Lectin histochemical characterisation of the porcine small intestine around weaning

The present study was undertaken to characterise the carbohydrate profile of the porcine small intestine using lectin histochemistry during the period from 3 days prior to weaning to 9 days post-weaning. A total of 56 piglets weaned at 4 weeks of age were included in the experiment. The most prominent changes in the glycosylation pattern were observed in the goblet cells. The highest lectin reactivity of the goblet cells in the crypts was observed 7 days post-weaning which suggests that the protective effect of the mucus layer against pathogenic bacteria is increasing during the postweaning period. The staining pattern of the apical membrane remained unchanged during the experimental period. This indicates that the glycosylation process in the goblet cells is rapidly inducible whereas changes in the glycosylation pattern of the apical membrane requires more time. The glycosylation pattern of both goblet cells and apical membrane differed between the positions of the small intestine. As glycoconjugates can act as attachment sites for microorganisms, these differences in the distribution of sugar residues may be one explanation for the site-specificity of certain pathogens.     

A dynamic model of protein digestion in the small intestine of pigs

A dynamic mathematical model of the digestion of proteins in the small intestine of pigs was developed. The model integrates current knowledge on the transit of digesta along the small intestine, endogenous secretions, digestion of proteins, and absorption of amino acids into a mechanistic representation of digestion. The main characteristics of the model are the following: the small intestine is divided into several segments of variable length but with equal digesta retention time; the rate of transfer of digesta between segments is based on the progression of myoelectric migration complexes; pancreatic and biliary secretions are poured into the first segment, whereas intestinal secretions enter all intestinal segments; protein hydrolysis is described by first-order equations; and an intestinal absorption capacity is used to estimate absorption of hydrolyzed protein. Simulation results are consistent with observed data, although more information is needed to represent reality more closely. The sensitivity analysis shows that parameters for protein hydrolysis largely determine protein digestibility. The absorption capacity of the small intestine limits the absorption of amino acids at the beginning of a meal and modulates the appearance of amino nitrogen in the portal vein. It also shows that amino acid absorption can be limiting to protein digestibility when large amounts of protein are eaten in a single daily meal. The model is useful in evaluating the dynamics of protein digestion and absorption of feedstuffs. The model can be used in evaluating protein digestion of different feedstuffs and feeding strategies.     

Small intestine and small colon neuropathy in equine dysautonomia (grass sickness)

The number of neurons in the coeliacomesenteric ganglia and the myenteric and submucosal plexuses of the jejunum, ileum and small colon, and the pathological changes induced in them, were studied in various types of equine dysautonomia. In all forms of dysautonomia, severe and extensive neuron loss and damage occurred in the ileum. In acute and subacute dysautonomia, jejunal neuron loss and damage were severe, but in chronic cases significantly less loss or damage occurred. The damage followed the same pattern in the small colon but it was always less obvious than in the jejunum. The distribution of the damage was uniform within a segment of the intestine. In fatal cases of dysautonomia, the clinical severity and duration of illness seems, in most instances, to be related to the amount of neuronal disruption occurring in the jejunum. Severe disruption results in acute/subacute dysautonomia, while milder damage leads to the chronic form.No case of dysautonomia was encountered in which enteric neuron loss and damage occurred without significant neuronal disruption also occurring in the coeliacomesenteric ganglia.Ileal neuronal damage and loss are not invariably worse than that in the jejunum, and the possible reasons for this, together with the relationship between neuronal damage and possible causes of dysautonomia, are discussed.Abbreviations H&E haematoxylin and eosin Deceased. Formerly of the Moredun Research Institute, 408 Gilmerton Road, Edinburgh, EH17 7JH, UK     

Xanthine oxidase formation during experimental ischemia of the equine small intestine.

We hypothesized that xanthine oxidase plays a role in the postischemic reperfusion injury in the equine small intestine. Under anesthesia, four horses and two ponies underwent ischemic strangulating obstructions of segments of the proximal jejunum, mid-jejunum and ileum. Prior to vascular occlusion, and at 1 h and 2 h of ischemia, full-thickness intestinal biopsies were collected for histopathological evaluation and for determination of combined xanthine dehydrogenase (XDH) plus xanthine oxidase (XO) activity, and XO activity alone. The level of XO activity was expressed in percentage according to the ratio of XO/(XDH + XO). We found a nearly threefold increase in the combined level of XDH plus XO activity from ileum to duodenum (p less than 0.04). However, the preischemic level of % XO activity did not vary significantly (p = 0.61) between segments of jejuno-ileum. Likewise, no significant difference was noted between intestinal segments after ischemia. Therefore, the data from all intestinal segments were pooled for each time and analyzed using Wilcoxon's signed rank test (one-tailed). Compared to the pre-ischemic level of % XO activity (median 27%), the % XO activity increased after 1 h of ischemia (median 37.0%), reaching statistical significance (p = 0.016). There were no statistical differences between the preischemic % XO activity and the % XO activity in non-ischemic bowel at the end of the anesthetic period. During ischemia, % XO activity increased, which lends credence to the importance of xanthine oxidase in previously-documented reperfusion injury in the equine small intestine.     

Dietary fructan carbohydrate increases amine production in the equine large intestine: implications for pasture-associated laminitis

Pasture-induced laminitis in the horse is associated with the overconsumption of fermentable carbohydrate, in the form of simple sugars, fructans, or starch. The fermentation of carbohydrate in the cecum and large intestine results in the production of lactic acid and other toxins or "laminitis trigger factors." Vasoactive amines have been suggested as possible initiating factors. The aim of this study was to feed a commercially available form of fructan carbohydrate (inulin, 3 g/kg of BW per day) to normal ponies and to ponies predisposed to laminitis, to mimic a change from a basal hay diet to lush spring-summer pasture. Five normal and 6 laminitis-prone, native-breed ponies were acclimated to a basal hay diet before the inclusion of inulin and chopped dried grass. Blood samples, fecal samples, and foot temperature measurements were taken throughout the study. Amines were measured in the feces and plasma by HPLC and liquid chromatography-mass spectrometry, respectively. The pH of the fecal samples decreased from 6.89 +/- 0.11 on the hay diet to a minimum of 6.18 +/- 0.11 with the addition of inulin (P <0.05). An increase was observed in the fecal concentrations of a number of amines, including tryptamine (2.5-fold increase, P <0.05) and tyramine (2-fold increase, P <0.05). No changes were noted in plasma amine concentrations or plasma D- or L-lactate, indicating that there may be a threshold of hindgut pH change before mucosal damage can result in the release of these factors into the circulation. No differences in pH or any of the measured compounds were observed between the group of normal ponies and those predisposed to laminitis. This indicates that differences in the intestinal microflora do not account for this predisposition. However, the results from this study indicate that moderate increases in dietary fructan carbohydrate can produce increases in bacterial fermentation products and other compounds in the large intestine, which may be relevant to the pathogenesis of acute laminitis in ponies on pasture.     

The significance of small vs large intestinal digestion of cereal grain and oilseed protein in the equine

Mature ponies fitted with permanent ileal cannulas were used in two 3×3 Latin square experiments to quantify prececal, postileal and total tract digestion of N. In trial 1, corn, oats and sorghum were each fed with coastal Bermuda grass hay in 75:25 ratios. Apparent prececal digestibilities were similar (P>.05) and averaged 46.6%. By-difference procedures were employed to calculate digestibility of the cereal grain N only and apparent prececal N digestibility averaged 57.1%. In trial 2, a basal corn and hay diet was supplemented with cottonseed meal and soybean meal. Apparent total tract N digestibilities were similar (P>.05) across treatments, and prececal digestibility averaged 45.6%. By-difference calculations were used to determine digestibility of SBM and CSM N alone. Apparent prececal digestibility of SBM was 52.5% and was lower (P<.05) than 81.2% for CSM. It is possible that inadequate or excessive heat treatment of SBM affected enzymatic digestion. True digestibility of total rations fed in trial 2 was estimated by linear regression of nitrogen digested on nitrogen intake or N presented to the large intestine. True N digestibility of diets containing SBM and CSM was 54.7% and 69.4%, respectively.     

Biomechanical characterisation of equine laryngeal cartilage

Reasons for performing the study: Upper airway obstruction is a common problem in the performance horse as the soft tissues of the larynx collapse into the airway, yet there is a paucity of information on biomechanical properties for the structural cartilage components. Objective: To measure the geometry and compressive mechanical properties of the hyaline cartilage to improve understanding of laryngeal function and morphology. Methods: A total of 11 larynges were harvested from Thoroughbred and Standardbred racehorses. During gross dissection, linear dimensions of the cricoid were obtained. From both the cricoid and arytenoid, specimens were cored to obtain 6 mm disc samples from 3 sites within the dorsal cricoid (caudal, middle and rostral) and 2 central sites in the arytenoids (inner, outer). The specimens were mechanically tested using radial confined compression to calculate the aggregate modulus and permeability of the tissue. The biomechanical data were analysed using a nested mixed effects model. Results: Geometrically, the cricoid has relatively straight walls compared to the morphology of human, ovine and canine larynges. There were significant observations of higher modulus with increasing age (0.13 MPa per year; P = 0.007) and stiffer cricoid cartilage (2.29 MPa) than the arytenoid cartilage (0.42 MPa; P<0.001), but no difference was observed between the left and right sides. Linear contrasts showed that the rostral aspect (2.51 MPa) of the cricoid was 20% stiffer than the caudal aspect (2.09 MPa; P = 0.025), with no difference between the arytenoid sites. Conclusions: The equine larynx is a well supported structure due to both the geometry and material properties of the cricoid cartilage. The hyaline structure is an order of magnitude higher in compressive modulus compared to the arytenoids and other hyaline‐composed tissues. Potential relevance: These characterisations are important to understand the biomechanics of laryngeal function and the mechanisms involved with surgical interventions.     

The characterisation of equine interleukin-1

Equine interleukin-1 has been produced from peripheral blood monocytes by stimulation with E. coli lipopolysaccharide. Sephacryl S200 gel filtration revealed a molecular weight of 17-18 kD. Chromatofocusing of the 17-18 kD peak identified four active fractions. Two major peaks were detected at pH 6.7 and pH 7, with smaller peaks at pH 6.3 and pH 5.9. The pI 7 molecule is probably the equine form of IL-1 beta.     

Nitrogen and amino acid absorption in the small intestine of growing pigs. 2. Apparent and true crude protein digestibility and amino acid absorption to the end of the small intestine

Studies of the apparent and true ileal digestibility of crude protein and the apparent and true amino acid (AA) absorption were carried out with 7 female pigs in the live weight range between 22 and 34 kg after ileorectostomy. Protein sources of varied quality with regard to the method of treatment (toasted) and untoasted soybean oilmeal) and to the AA composition (vital gluten or vital gluten + L-lysine X HCl) were used. Distinct differences regarding the apparent ileal digestibility of crude protein and the apparent absorption of amino acids were discovered between the protein sources, the cause of which is to be seen in the different amounts of endogenous N. The true ileal digestibility of crude protein and amino acid absorption were partly distinctly higher than the values of the apparent ileal digestibility of crude protein and AA absorption. On the whole one can say that the results of the investigation gained by means of ileorectostomy can be compared to the values gained from cannulated animals. Thus the experimental method used is suitable for studying the absorption process to the end of the small intestine of pigs.     

北京雏鸭小肠消化酶活性变化规律研究

北京鸭成熟早,生长发育快,育肥性能好,肉质细嫩,味道鲜美,是我国著名的肉用鸭品种.近年来,关于通过添加外源性消化酶以弥补家禽体内消化酶的不足、降低肠道食糜黏度,从而提高饲料利用率、增强机体代谢水平和免疫力以促进动物生长方面的研究较多[1],但对北京雏鸭内源酶的变化规律未见报道.     

Molecular characterisation of equine group A rotavirus, Nasuno, isolated in Tochigi Prefecture, Japan

In this study, equine group A rotavirus (RV-A), Nasuno, isolated from foal diarrhoea in Tochigi Prefecture, Japan was characterised genetically by sequence analysis of the genome segments encoding VP4 and VP7. The nucleotide and deduced amino acid sequences revealed high homology with P[12] RV-As (94.0-99.3% and 94.9-99.4%) and G3 RV-As (86.9-99.5% and 91.1-99.4%). Nasuno was also classified into P[12] and G3 in the phylogenetic analysis of the nucleotide sequences of the genome segments encoding VP4 and VP7.