Effects of dietary change and rotavirus infection on small intestinal structure and function in gnotobiotic piglets

The combined effects of weaning and rotavirus infection on small intestinal structure and function and on growth rate were studied in 28 gnotobiotic piglets. There was little damage by rotavirus to the proximal small intestine, some damage to the mid small intestine and relatively severe damage to the distal small intestine; villi were stunted, crypts lengthened and activities of all brush border enzymes decreased. The damage was short-lived despite the synchronisation of rotavirus infection with simulated weaning. There was no evidence of persistent damage to the small intestine and growth rate was unaffected.     

The gastrointestinal mucosa in young milk-fed calves. A scanning electron and light microscopic investigation

Gastrointestinal segments from 4 healthy, 17-, 21-, 22- and 23-day-old calves fed on whole cow’s milk were examined. Scanning electron microscopy showed that the anterior duodenum had short villi varying in shape from leaf-shaped to nodular; the middle duodenum had broad, tongue-shaped villi and the anterior, middle, and parts of the posterior jejunum had slender, finger-shaped or leaf-shaped villi. The villi of the mucosa covering Peyer’s patches in the posterior jejunum were short and either conical or tongue-shaped; there were also small “pseudovilli” caused by bulges in the lymphoid tissue. Morphometry showed that the villi were longer in the anterior jejunum than in the duodenum and the posterior parts of the jejunum (P < 0.005). Morphologically fat absorption was most heavy in the anterior third of the small intestine. Moderate amounts of fat were also found in the epithelium of the posterior jejunum and of the abomasum. Large fat droplets were seen in apical duodenal enterocytes, in contrast to the small epithelial droplets in other areas with fat absorption. Nile blue staining indicated that the fat in the large droplets was esterified.     

Decreased activity of brush border membrane-bound digestive enzymes in small intestines from pigs experimentally infected with porcine epidemic diarrhea virus

Brush border membrane-bound digestive enzymes such as disaccharidases (lactase, sucrase, and maltase), leucine aminopeptidase N, and alkaline phosphatase were measured in jejunum from pigs experimentally infected with porcine epidemic diarrhea virus (PEDV). Three piglets from the infected and control groups were euthanized by electrocution and subjected to necropsy at 24, 36, 48, 60, and 72 hours post-inoculation (hpi). The infection of PEDV to jejunum resulted in significant decreases in brush border membrane-bound digestive enzymes such as disaccharidases (lactase, sucrase, and maltase), leucine aminopeptidase N, and alkaline phosphatase. PEDV replication results in massive destruction of villous enterocytes leading to a marked reduction of intestinal epithelial surface and brush border membrane-bound digestive enzyme activity. Reduced enzymatic activity and villous atrophy in the small intestine is thought to result in a maldigestive and malabsorptive diarrhea.     

Cell differentiation in intestinal adenomatosis of pigs studied by histochemistry of laminin and enzymes of epithelial and subepithelial tissue

The distribution of enzymes and laminin was examined in ileal tissue from pigs suffering from intestinal adenomatosis to reveal the nature of the lesion. A disruption of the normal and specific pattern of distribution was found. Thus, the normal ileal epithelium was characterised by brush border enzymes: alkaline phosphatase, magnesium-dependent adenosine triphosphatase (Mg-ATPase), fluoride resistant acid phosphatase and 5'-nucleotidase; enzymes of the basolateral border: Mg-ATPase; and cytoplasmic enzymes: beta-glucuronidase, non-specific esterase and acid phosphatase. Subepithelial fibroblasts seemed to be characterised by 5'-nucleotidase. Laminin was present as a continuous band under the surface and crypt epithelium, somewhat thicker in the former. In contrast, the branching proliferating crypts of intestinal adenomatosis largely lacked enzymes characteristic of both villus and crypt cells. Reactions for the subepithelial components, laminin and fibroblasts were also reduced. The deficient differentiation of the epithelial as well as subepithelial components in porcine intestinal adenomatosis distinguish the condition from crypt hyperplasia and indicate an adenoma-like character.     

Development of wheat-sensitive enteropathy in Irish Setters: biochemical changes

Biochemical changes in the small intestine during development of naturally acquired wheat-sensitive enteropathy of Irish Setters were investigated. To distinguish primary biochemical abnormalities from secondary effects of intestinal damage, progeny of affected dogs reared on a normal wheat-containing diet were compared with their own littermates reared on a cereal-free diet and with age-matched clinically normal Irish Setters fed the same wheat-containing diet. Peroral jejunal biopsy specimens were sequentially obtained between weaning and 1 year of age; specific activity and reorientating sucrose density-gradient distribution of organelle marker enzymes were determined. Major primary biochemical abnormalities were not detected in affected progeny. In affected dogs fed wheat, there was a selective, but secondary, loss of the brush border alkaline phosphatase and aminopeptidase N activities. This loss was associated with the development of partial villus atrophy, but represented a specific effect of dietary wheat on the brush border, not merely a nonspecific effect of mucosal damage, because other brush border enzymes, including disaccharidases, were not similarly affected. Increased soluble activities of lysosomal and peroxisomal marker enzymes late in the disease process may represent alterations in these 2 organelles as a secondary consequence of mucosal damage.     

Ultrastructural changes in the small intestine of neonatal calves with enteric colibacillosis

The small intestines of calves inoculated orally with the enteropathogenic strain of Escherichia coli 0101:K'B41',K99 were examined by electron microscopy at 3, 6, 12, 16, 21, 36, 69, 70 and 72 hours after inoculation. The challenge organism adhered to the mucosa of the distal small intestine from six hours post-inoculation. Bacteria were separated from the microvillous brush border by a gap of 200 to 300 nm in which bacterial fimbriae and the microvillous glycocalyx were seen. Bacteria never were found in epithelial cells but were present in macrophages in the lamina propria from 12 hours. At three and six hours, cytopathic changes were not seen in the small intestine, but from 12 hours epithelial cells on affected villi had blunt and thick microvilli and contained cytoplasmic inclusions. Epithelial cells were seen frequently in the process of extrusion from the villi, either singly, in small groups, or as ribbons of cells. Intervillous bridges, characteristic of villous fusion, were seen frequently from 69 hours.     

Simultaneous isolation and characterization of brush border and basolateral membrane vesicles from bovine small intestine

Purified brush border and basolateral membranes were isolated from homogenized intestinal enterocytes of Holstein steers by divalent cation precipitation followed by differential and sucrose density gradient centrifugation. Alkaline phosphatase and Na/K adenosine triphosphatase served as marker enzymes for the brush border and basolateral membranes, respectively. The brush border and basolateral membrane fractions were enriched 5.1- and 10.1-fold, respectively, over the cellular homogenate. Electron micrographs, obtained with transmission electron microscopy, confirmed the vesicular nature of the membranes and revealed that basolateral membrane vesicles generally were smaller and more irregular in shape than brush border membrane vesicles. The vesicular nature of isolated membrane preparations was confirmed with osmotic activity experiments. Enrichment of brush border and basolateral membrane fractions compared to the initial homogenate and the vesicular configuration of both preparations indicate that the isolated brush border and basolateral membrane preparations were suitable models for evaluating nutrient transport properties of bovine small intestine. The number of transport experiments possible per animal using the membrane vesicle technique is many times more efficient than some other in vitro techniques (i.e., intestinal rings or everted sacs).     

Über die pränatale Entwicklung der Lamina epithelialis in der Phase der Vakuolisierung villärer Epithelzellen. Untersuchungen am Darm des Rindes (Bos primigenius taurus L., 1758)

The development of the bovine small intestine was examined in 39 embryos and fetuses by light microscopic and transmission electron microscopic methods. Special reference was paid to the histogenesis of the ephithelium. In contrast to the duodenum the epithelium of jejunum and ileum undergoes a degeneration by vacuolation of its villous epithelial cells. The demonstration of the acid phosphatase activity of these vacuoles shows their lysosomal character. This degenerative process of the small intestinal epithelium is also known in large intestine where it leads to the destruction of the intestinal villi. Both seem to be part of a ‘principle of construction of the intestine of the vertebrates’(Wille, 1984).     

实验性流行性腹泻仔猪小肠粘膜上皮细胞及肠系膜淋巴结的超微结构

给8头生后3d的哺乳仔猪经口感染猪流行性腹泻病毒(PEDV)“吉”毒株,于感染后18、30、45和96h各扑杀2头,以透射电镜和扫描电镜观察了小肠粘膜上皮细胞及肠系膜淋巴结的超微结构。结果表明,小肠上皮细胞的病变因感染时间不同而有明显差异。上皮细胞的脱落和残留上皮细胞超微结构的破坏,以感染后30h最严重,病毒在这些上皮细胞内的增殖最显著。感染后45h,见有大量新生上皮细胞修补损伤的肠绒毛。感染后96h,小肠绒毛短缩、粗大乃至发生融合。实验仔猪肠系膜淋巴结内巨噬细胞和淋巴细胞的超微结构均遭到破坏,在巨噬细胞内见有PED冠状病毒粒子。     

Chick small intestine brush border contains lipase and phospholipase activity.

1. Uptake of triglycerides and phosphoglycerides was demonstrated in ligated washed chick intestinal segments in situ. Mucosal uptake was observed both in the jejunum and ileum of chicks and was not accompanied by significant luminal hydrolysis of acylglycerides. 2. Brush border membrane vesicles were prepared and lipase and phospholipase activities were demonstrated. These activities were enriched in parallel to the increase in activity of maltase and gamma-glutamyl transpeptidase in all small intestinal areas. 3. In order to determine if the pancreatic and brush border membrane lipolytic activities were similar different properties of these activities were examined. The relative activities of pancreatic and brush border membranes towards triolein and phosphatidylcholine differed. The pH-triolein activity curve and the use of different additives showed some diverging effects between the activities. These differences were not, however, sufficient to conclude that the brush border activity towards triolein is distinct from that of pancreatic lipase. 4. Lipolytic activity in the brush border may play a role in acylglyceride digestion, uptake and transport in the small intestine of the chick.     

Histochemistry of acid phosphatase in small intestine mucosa in experimental coccidiosis in suckling piglets]

The activity of acid phosphatase (phosphohydrolase of orthophosphate monoesters; EC. 3.1.3.2) was evaluated densitometrically in the mucosa of duodenum, jejunum and ileum of 22 conventional piglets which were experimentally infected by oocysts of the coccidiae Isospora suis (infection dose of 200,000 oocysts) on day one after parturition (DAP). The activity of the studied hydrolase was investigated in the infected piglets during days two to ten after infection (DAI) in the intestinal mucosa (enterocytes) and in goblet cells. The density of the reaction product of acid phosphatase was simultaneously determined in the same mucosal cells of different sections of the small intestine in five control conventional piglets at the age of 2-14 days. In the small intestine mucosa of control piglets the activity of acid phosphatase was demonstrated to be located especially in the supranuclear zone of enterocytes. As for goblet cells, the reaction product of acid phosphatase is distributed in all zones (supra-, para-, infranuclear zones); the lowest density of this enzyme was found in the infranuclear zone. The activity of acid phosphatase is also localized in intestinal crypts: in their cells the enzyme concentration is decreasing from duodenum to caudal sections. Important changes were revealed, in comparison with the control data, in the development of the activity of acid phosphatase in the intestinal mucosa cells in the experimentally infected piglets. In the period of investigation (DAI 2-10) there were two stages of the development of the density of the enzyme reaction product. The first stage can be characterized by an increase, the other by a decrease in the level of acid phosphatase activity. Enterocytes are influenced in both stages, but the decrease in the density of the reaction product of acid phosphatase was observed only in absorption cells, and not in goblet cells. The increase in the activity of acid phosphatase occurs in the periods of DAI 4 and 9-10. Enzymatic deviations occur mainly in the absorption cells of the mucosa of duodenum and middle jejunum; in the cells of posterior jejunum and ileum an increase in the density of the reaction product of acid phosphatase was also demonstrated, but at the lower quantitative level (especially on DAI 4). The decrease in the activity of acid phosphatase has a protracted development and it takes place on DAI 5 to 8.(ABSTRACT TRUNCATED AT 400 WORDS)     

Intestinal enzyme activity in lambs chronically infected with Trichostrongylus colubriformis: effect of anthelmintic treatment

After twenty weeks of continuous dosing with Trichostrongylus colubriformis larvae substantial, but declining, numbers of worms had persisted in most of the lambs examined, although there were wide inter-individual variations. Mucosal lesions were found in the proximal small intestines of all the infected animals, their severity being directly related to worm burden. Representative brush border enzyme activities analysed in intestinal mucosal extracts from the same lambs showed differing responses. Alkaline phosphatase and glycyl-L-leucine dipeptidase were significantly depleted, whereas maltase activity was only marginally reduced, and leucine aminopeptidase activity was normal. Mucosal acetylcholinesterase activity was significantly elevated in the parasitised animals and, interestingly in view of the postulated role of this enzyme in nematode pathogenicity, the level of activity was directly correlated with individual worm burdens. Intestinal trypsin and chymotrypsin activities were unaffected and the level of superoxide dismutase, an enzyme associated with the inflammatory response, was normal. There were also no consistent changes in the mucosal activities of several enzymes including lactic dehydrogenase, creatine phosphokinase, aldolase, and glutamic oxaloacetate transaminase, whose leakage from damaged or necrotic tissues has been well defined in terms of the concomitant increase in their activity in the circulation. Lambs treated orally with fenbendazole five and/or ten weeks before slaughter either in the presence or absence of continued larval intake, had negligible worm burdens, and showed little evidence of intestinal damage at post mortem. Brush border enzyme levels, with the exception of alkaline phosphatase and, in two cases dipeptidase, were normal in these animals. The activity of alkaline phosphatase was approximately double that in the continuously infected, untreated lambs, but remained markedly lower than in the uninfected controls. The activities of the other enzymes studied, including acetylcholinesterase, were within the control range. In summary, in chronic trichostrongylosis even relatively low nematode burdens were associated with marked pathological and biochemical damage in the intestine with both lesion severity and mucosal acetylcholinesterase activity being directly related to worm numbers. Although morphological integrity was completely restored after anthelmintic treatment, the persistent low activity of brush border alkaline phosphatase coupled with the enzymological findings in untreated, infected animals suggests that recovery of the full functional capability of the intestinal mucosa may take longer.     

Changes in intestinal villi, cell area and intracellular autophagic vacuoles related to intestinal function in chickens

1. Morphological changes in the intestinal villi, cell area and cell mitosis number in the duodenal epithelial cells were compared in cockerels fasted for 1, 2 and 3 d, and also when refed for 1 and 2 d after 3 d of fasting, to demonstrate whether these morphological changes are related to intestinal function. Alterations in the fine structure of vacuoles in epithelial cells were also examined in each group to investigate whether the vacuolar changes are associated with these morphological changes, and to obtain an index for judging the nutritional condition of the chicken intestine. 2. Fasting induced decreases in villus height, cell area and cell mitosis number, which recovered rapidly after refeeding, suggesting that these parameters are related to changes in intestinal function and may be useful for assessing intestinal function. 3. At 1 d of fasting, small electron-dense bodies appeared in the absorptive epithelial cells, some of them fusing with each other. As the fasting period increased, these small bodies developed to moderate-sized nascent autophagic vacuoles containing various kinds of electron-dense contents and finally became large autophagic vacuoles with electron-lucent contents. Some vacuoles showed positive acid phosphatase reactions, which indicated that they were lysosomal autophagic vacuoles containing hydrolytic enzymes. 4. After 1 d of refeeding the large autophagic vacuoles seen after 3 d fasting rapidly decreased to the small electron-dense bodies seen after 1 d of fasting. 5. These findings suggest that intestinal epithelial cells have the ability to digest their own cell components to supply nutrients during fasting by means of lysosomal active autophagic transport mechanisms: after refeeding, the epithelial cells return to the absorption of nutrients. 6.The present results demonstrate that the autophagic vacuolar changes are correlated with changes in intestinal villus height, cell area and cell mitosis number induced by fasting and refeeding; this indicates that autophagic vacuoles are a useful index of the nutritional condition of chicken intestine. The greater the number of electron-lucent vacuoles there are in the duodenal absorptive cells, the lower the nutritional condition of the chicken intestine.     

Histochemical analyses of digestive enzymes in the intestine of adult large-scaled gurnard (lepidotrigla cavillone, lacepède, 1801)

Localization and activity levels of the following digestive enzymes in the intestine of free-living large-scaled gurnard were determined: non-specific esterase, alkaline and acid phosphatase as well as aminopeptidase. Enzymatic activity of the four enzymes was confirmed in all intestine parts but with different distribution through the enterocytes and varying in diverse intensity according to intestine part. This research is part of a broader research project on the biology of economically important fish from the Adriatic Sea. Our study reveals that in the large-scaled gurnard, the middle and posterior intestinal segments play the major role in digestion and absorption of proteins, whereas all parts of the intestine participate in lipid absorption and intracellular digestion. The high protein and lipid content in the diet of the large-scaled gurnard is most likely responsible for high activities of esterase, alkaline phosphatase and aminopeptidase, as they are involved in digestion and absorption of proteins and lipids.     

Enzymes of isolated brush border membrane of Cotugnia digonopora, and their insensitivity to anthelmintics in vitro

Purified brush border membrane of Cotugnia digonopora showed the presence of a number of phosphohydrolases. Among these, alkaline phosphatase was extremely active. Other enzymes such as glucose-6-phosphatase, fructose-1,6-diphosphatase, cAMP-phosphodiesterase, 5'-nucleotidase and adenosine-triphosphatase were also active. Observations were made on the activities of various ATPases; whereas the enzyme was activated by Ca++ and Mg++ in an additive manner, its sensitivity to ouabain was negligible. Furthermore, in the presence of EDTA the enzyme activity was quite significant. The treatment of isolated brush border membrane with mebendazole, niclosamide and praziquantel in vitro did not alter the activity of these enzymes. However, treatment of intact worms drastically affected the integrity of the membrane.     

Über die frühfetale Entwicklung der Dünndarm-Mukosa des Rindes (Bos primigenius taurus).

The development of the bovine small intestine was examined in 24 embryos and fetuses by light microscopic, scanning and transmission electron microscopic methods. Special reference was paid to the genesis of the epithelium and particularly of the villi intestinales. The primitive intestine consists of one layer of epithelial cells surrounded by mesenchym and tunica serosa. The fetal intestine (up to the 24th week of gestation) shows all the morphologic structures of the adult. In small intestine the development of cryptae and villi intestinales starts before the 7th week of gestation and progresses with a proximo-distal gradient. Epithelial proliferation that gives rise to primary epithelial villi makes epithelium become temporarily stratified. Finger-like secondary villi develop by proliferation of the mesenchym. In addition to this process mucosal folds occur in duodenum giving rise to villi by segmentation. At the same time the differentiation of epithelium starts. The fetal small intestine, like many other fetal tissues displays masses of glycogen.     

高脂饲料对细鳞鱼稚鱼刷状缘膜水解酶发育的影响

试验评价了蛋白质含量为48%时,脂肪含量分别为15%、20%、25%、30%的饲料对开口后20～60 d的细鳞鱼稚鱼肠道水解酶-γ-谷氨酰肽转移酶、碱性磷酸酶活性发育的影响。结果表明,适度的饲料脂肪含量可以提高水解酶活性,并强于生物饵料,促进了肠道对营养物质的消化。饲料脂肪为20%～30%的水平时,水解酶的活性相对较高。     

INTESTINAL CRYPTOSPORIDIOSIS IN A KID GOAT

SUMMARY: A 2-week-old Angora kid goat developed diarrhoea and was treated with parenterally administered antibiotics and saline. The kid rapidly became moribund and died within 6 hours of the first signs of illness. No bacterial or viral agents were detected in the intestinal content. On histological examination large numbers of small haematoxylin-staining bodies were detected associated with the brush border of small intestinal enterocytes. Associated with these bodies there was variable shortening of intestinal villi, some congestion and focal leucocyte infiltration in the lamina propira of the villi and focal epithelial erosions, sloughing or cell transition to more immature enterocyte forms. The location and features revealed by light microscopy and scanning and transmission electron microscopy indicate these bodies are cryptoporidia.