Ultrasonographic imaging of abomasal milk clotting and abomasal diameter in healthy and diarrheic calves.

In case of diarrhea calves are treated with oral rehydration solutions (ORS), which are known to increase abomasal pH and inhibit milk clotting in vitro. Nevertheless, recent studies have shown that ORS with HCO3(-) ≤ 62 mmol/L do not interfere with abomasal milk clotting in healthy calves. However, in diarrheic calves, feeding ORS and milk simultaneously may disturb abomasal curd formation and exacerbate diarrhea due to faster abomasal passage of ingesta. Therefore, the aim of the present study was to ultrasonographically examine abomasal milk clotting and diameter after feeding milk and milk replacer (MR) with and without ORS to healthy and diarrheic calves. Abomasal curd formation and diameter in healthy and diarrheic calves were ultrasonographically imaged before and after feeding milk, MR and ORS prepared in milk or MR. Feeding mixtures of milk or MR with ORS did not cause any remarkable differences in the ultrasonographic images of abomasal content. Moreover, abomasal milk clotting was not disturbed due to diarrhea. Statistically significant differences of abomasal diameter after feeding between healthy and diarrheic calves indicated that abomasal emptying is delayed in diarrheic calves. Hence, further studies are needed to determine reasons for decelerated abomasal passage in calves suffering from diarrhea.

Histomorphometric adaptation of yak (Bos grunniens) abomasum to the Qinghai-Tibetan plateau environment / Adaptación histomorfométrica del abomaso de yak (Yos grunniens) al medio ambiente de la meseta tibetana Qinghai

Six abomasums of yaks (Bosgrunniens) were studied with gross dissection and histological methods. It was found that the mucosa of the yak abomasum was covered with simple columnar epithelium. There were lots of spiral folds (10) in the fundic glandular area. The developed membrane of lamina propria was occupied by high density glands. According to the morphological characteristics of the glands, the abomasum was divided into the cardiac, gastric and pyloric glands. Cardiac glands were curved tubular glands with the intumescent bottom and small glandular cavity. Fundic glands were simple tubular glands or branched tubular glands, where the chief, parietal and mucous neck cells can be observed clearly. Pyloric glands were curled tubular glands, the closer to the deep of the lamina propria, the more obvious the glands curl. Staining of glycoconjugate revealed that the mucosal epithelium of the cardiac gastric and pyloric glands and gastric pits epithelium mainly secreted neutral glycoconjugate, but other portions of cardiac and gastric glands secreted mixed and acid glycoconjugate respectively. By Gordon-Sweet's reticular fiber staining, it was found that the mucous neck cells possessed the characteristic of argyrophilic phenomenon. There was a large number of argyrophilic granules in the supranuclear cytoplasm in contrast with the chief cells. Furthermore, there were isolated lymphoid nodules and diffuse lymphoid tissue in the abomasum glands, especially in corpus abomasi. Grimelius silver staining showed that the argyrophil cells were located in the glandular epithelium and lamina propria of glands, which can also be observed in connective tissue. These endocrine cells dispersed individually in epithelial cells, occasionally in 3­5 cell groups. Therefore, the yaks were grazed throughout the year on diverse natural grassland and had evolved morphological characteristics of the abomasum enabling them to consume a wide variety of plant species, thereby better adapting them to harsh plateau environment.

Seis abomaso yak (Bosgrunniens) fueron estudiados con disección y métodos histológicos. Se encontró que la mucosa del abomaso yak estaba cubierta de epitelio columnar simple. Se observaron pliegues en espiral (10) en la zona glandular fúndica. La membrana desarrollada de la lámina propia contenía glándulas de alta densidad. De acuerdo con las características morfológicas de las glándulas, el abomaso se dividió en las glándulas cardíacas, gástricas y pilórica. Las glándulas cardíacas se curvan en glándulas tubulares con la parte inferior intumescente y una pequeña cavidad glandular. Las glándulas fúndicas eran glándulas tubulares simples o glándulas tubulares ramificadas, donde se pueden observar con claridad las células principales, parietales y mucosas del cuello. Las glándulas pilóricas fueron glándulas tubulares curvadas, cuanto más cercanas a la lámina propia, más evidente fue su forma ondulada. La tinción glucoconjugada reveló que el epitelio de la mucosa de las glándulas gástricas cardiacas, pilóricas y el epitelio de las fosas gástricas secretaron principalmente un glucoconjugado neutro, pero otras porciones cárdicas y de las glándulas gástricas secretaron un glucoconjugado mixto y ácido, respectivamente. A la tinción de fibras reticulares, se encontró que las células mucosas del cuello poseían características argirófilas. Se observó un gran número de gránulos en el citoplasma supranuclear en contraste con las células principales. Además, no fueron aislados los nódulos linfoides y presentaba tejido linfoide difuso en las glándulas de abomaso, especialmente en el cuerpo del abomaso. La tinción Gordon Sweet indicó que las células argirofílicas se localizaron en el epitelio y lámina propia glandular, lo que también se observó en el tejido conectivo. Estas células endocrinas se dispersan individualmente en las células epiteliales, de vez en cuando en grupos celulares de 3-5. De esta forma, los yak pastorean durante todo el año, en diversos pastizales naturales, y han evolucionado sus características morfológicas que les permiten consumir una amplia variedad de especies de plantas, con lo que se adaptan mejor a las condiciones inhóspitas.

Embryonal carcinoma in the abdominal cavity of a male calf.

An embryonal carcinoma was diagnosed in the abdominal cavity of a 55-day-old male calf. Macroscopically, a large volume of ascitic fluid was observed along with white to yellowish-white masses of various sizes densely located on the abdominal wall and the surface of abdominal organs. There was an absence of testes in the scrotum. Histologically, the tumor cells were polygonal, and the tumor was mostly composed of sheets of densely packed solid patterns with occasional papillary and tubular structures. Cell nuclei were variable in size, and cellular mitotic rate was high. Immunohistochemically, tumor cells were positive for alpha-fetoprotein, placental alkaline phosphatase, cytokeratin, and carcinoembryonic antigen. Ultrastructurally, the tumor cells had large nuclei, extensive rough endoplasmic reticulum, and small numbers of mitochondria. Microvillus-like structures and desmosomes were occasionally observed. From lectin histochemical examination, the tumor cells were positive for concanavalin A, wheat germ agglutinin, peanut agglutinin, Dolichos biflorus agglutinin, soybean agglutinin, Griffonia simplicifolia I, and Bauhinia purpurea, and negative for Ulex europaeus agglutinin I. Results of histopathological, immunohistochemical, and ultrastructural examinations of the tumor were similar to those obtained for human embryonal carcinoma.

Eosinophil interactions with Haemonchus contortus larvae in the ovine gastrointestinal tract.

Sheep were immunized by weekly oral infections with Haemonchus contortus for 9 weeks followed by anthelmintic treatment. They were challenged either 9 or 22 weeks later with PBS (sham controls) or one million exsheathed L3 surgically injected in the abomasum, and killed 24 h or 48 h later. Sheep challenged 9 weeks after immunization displayed varying degrees of tissue eosinophilia that showed a significant inverse relationship with the number of intra-epithelial mast cells (globule leucocytes). Close association of eosinophils with tissue larvae was observed mainly in the gastric pits (24 h) or on the mucosal surface (48 h). All L3-challenged sheep in this group had detectable globule leucocytes and tissue IL-4 mRNA, as measured by Southern blot RT-PCR. In contrast, sheep challenged 22 weeks after immunization had no detectable globule leucocytes or IL-4 mRNA and although they exhibited consistent tissue eosinophilia, eosinophils were not closely associated with tissue larvae. Scanning and transmission electron microscopy of sheep sensitized and rested for 9 weeks before challenge showed that L3 surrounded by eosinophils were at varying stages of damage and structural collapse. These studies strongly indicate that eosinophils can damage and probably kill gastrointestinal nematode larvae in vivo. In addition, they also suggest that effective killing by tissue eosinophils may depend on other microenvironmental factors such as intra-epithelial mast cells and IL-4.

A macro- and light microscopical study of the pathology of Calicophoron microbothrium infection in experimentally infected cattle.

Twelve Tuli weaner steers aged 1 year were randomly subdivided into three groups of four animals and infected with different doses of Calicophoron microbothrium metacercariae. Each animal in Group I received a low dose (LD) of 5 000 metacercariae, Group II a medium dose (MD) of 15 000 metacercariae, Group III a high dose (HD) of 25 000 metacercariae and one additional animal was kept as an uninfected control (C). After infection, one animal from each group was slaughtered on Day 28, 42, 56 and 84 post infection (pi) and samples from the ileum, jejunum, duodenum, abomasum and the rumen were collected for histopathological and cytological examination. On Day 28 pi, the gross pathological lesions observed in the duodenum of the LD and the MD animals were similar and comprised duodenal thickening, corrugation, hyperaemia, petechiation and ulceration. In the HD animal the duodenal lesions were similar but more severe. The abomasal folds were severely oedematous in the MD group and nearly occluded the abomasal lumen. Moderate oedema of the abomasal folds was also present in the LD and HD animals. The gross pathological lesions regressed in all the infected groups with increasing age of infection and had disappeared completely by Day 56 pi. On Day 28 pi the histopathological lesions in the duodenum and jejunum of the LD and MD groups were similar, comprising subtotal villous atrophy, hyperplasia of Brunner's glands and Peyer's patches and moderate infiltration of eosinophils, mast cells and a few globule leukocytes, basophils and lymphocytes in the lamina propria. The HD group had total villous atrophy, severe hyperplasia and cystic dilatation of Brunner's glands, which had expanded to cover the entire submucosa. On Day 42 pi the histopathological lesions were still present in the MD and the HD groups comprising subtotal villous atrophy and hyperplasia of Brunner's glands. Heavy infiltrations of eosinophils, moderate amounts of mast cells and a few basophils, globule leukocytes and lymphocytes were still present in the lamina propria of all three groups. On Day 56 pi, a few glands were still cystic in the MD and the HD groups. Moderate cell infiltrations were still present in the lamina propria of all the three groups and by Day 84 pi complete regeneration had occurred in all animals.

Light and electron microscopic investigation of the lectin-binding pattern in the oxyntic gland region of bovine abomasum.

For the first time the expression of glycoconjugate residues in the oxyntic gland region of bovine abomasum has been investigated by means of lectin histochemistry. For light microscopic investigations, a battery of ten lectins, Con A, PSA, UEA I, WGA, LEA, SNA, RCA120, MPA, DBA and SBA was used. For electron microscopic examinations, WGA and RCA120 were utilized. The staining pattern of the lectins in all exocrine cell types of the oxyntic gland region is described. Compared to the results of monogastric species our study reveals some similarities, but just as many differences in the composition of glycoconjugate residues in bovine exocrine cell types. Typical for surface mucous cells is the amount of L-fucose, N-acetyl glucosamine residues and Galbeta1, 4GlcNAc sequences in the secretory granules. SNA could serve as a marker for surface mucous cells, because this lectin exclusively stains the plasma membrane and the secretory granules of surface mucous cells and the extracellular mucus. L-fucose and N-acetyl glucosamine are typical for the secretory granules of mucous neck cells. In addition, the secretory granules show the highest amount of N-acetyl galactosamine residues of all exocrine cells, so that DBA and SBA are recommended as marker lectins for mucous neck cells. Most lectins strongly stain the intracellular membrane system of oxyntic cells. The cocktail of glycoconjugates in the vicinity of the HCI production site provide protection against chemical injury. In chief cells only the apical plasma membrane is more or less labeled with all lectins apart from SNA. Specific marker lectins for oxyntic cells or chief cells of the bovine have not been characterized.

Evidence for a parasite-mediated inhibition of abomasal acid secretion in sheep infected with Ostertagia leptospicularis.

The acid secretory capacity of the abomasal mucosa was studied in sheep experimentally infected with Ostertagia leptospicularis. The acidity of the abomasal contents, permanently recorded by a pH probe located inside the abomasum, decreased markedly to mean levels between pH 5 and 6. Subcutaneous administration of histamine or carbachol successfully stimulated acid secretion (pH 3.4). The results indicate that the abomasal mucosa harboured a population of functional parietal cells which were also identified immunohistochemically (H(+)/K(+)-ATPase). Ultrastructural investigation before stimulation revealed that the majority of these cells was in a resting state. Despite high serum gastrin levels, the acid secretion was blocked either at the level of the parietal cell or the enterochromaffin-like cell by an unknown factor, possibly mediated by the parasites. This is the first report of a parietal cell dysfunction associated with a nematode infection in the abomasum. It is suggested that the parasites induce changes in their environment which favour their survival and/or increase their reproduction.

Electron-microscopic detection of cytoskeletal actin in the epithelial cells of bovine abomasum.

The fine structural distribution of cytoskeletal actin was investigated in the oxyntic gland region and in the pyloric gland region of bovine abomasal mucosa using immunoelectron methods. In all exocrine cell types of the gastric epithelium, actin filaments exhibit a polarized pattern of distribution. In the members of the mucus- and protein-synthesizing cell line, the actin filaments form a prominent barrier to the exocytosis of secretory granules underneath the apical membrane. Therefore, actin is supposed to be involved in the control of exocytosis. The basolateral subplasmalemmal cortex of the exocrine cells, however, shows weaker labeling for actin filaments. In this position actin might be responsible for the movement of epithelial cells along the length of the gastric gland during physiological cell renewal and for the maintenance of the integrity of the epithelial unit and of cell polarization. In parietal cells of bovine abomasum, actin filaments are, in addition, associated with the intracellular canaliculus, but not with the tubulovesicular compartment. We assume that actin filaments reorganize the canalicular membrane after membrane translocations in the course of acid secretion. For the first time, actin filaments were documented in the rare brush cell type of bovine abomasum, where they do not form a terminal web beneath the apical membrane and therefore point out the nonexocrine function of the brush cell type. In all polarized cells of the bovine abomasum, actin filaments underlay the two apical members of the junctional complex, the tight junction and the adherens junction, where they might be involved in the regulation of cell adhesion, cell motility and cell shape determination.

Ultrastructure of the zymogenic-cell lineage in abomasal mucosa of adult cattle.

The ultrastructural differentiation and maturation of the neck cells and the zymogenic cells during physiological cell renewal were investigated in the abomasal oxyntic-gland region of cattle. Immature neck cells of the distal isthmus and proximal neck exhibit transitional morphology to the predominantly mucous isthmus cells. Neck cells confined to the glandular neck are characterized by bipartite peptic-cored mucous secretory granules. In a proximal-distal gradient along the neck, a progressive increase in the peptic granular component and concomitant reduction in mucous components paralleled by proliferation of the rough endoplasmic reticulum creates pre-zymogenic cells in the proximal glandular base. These, in turn, give rise to mature zymogenic cells with pure peptic secretory granules and typical zymogenic cell morphology. In the depth of the gland, older degenerative zymogenic cells are found. Variations in size and number of the zymogenic granules point to different secretory activities of the mature zymogenic-cell population of the glandular base. These results favour the conception of a zymogenic-cell lineage arising within the isthmus and passing through different developmental stages, including neck cells, during their migration down the gland.

[The differentiation of the surface mucous-cell line in the abomasum of the adult cow]. / Die Differenzierung der Oberflächenzellinie im Labmagen adulter Rinder.

The differentiation of the surface mucous-cell lineage during physiological cell renewal was investigated using light and electron microscopy in the abomasal mucosa of adult cattle. The surface mucous cells constitute a morphologically and functionally heterogeneous population, whose members correspond to different developmental stages, OFFanged in a distoproximal gradient from the depth of the pit towards the free luminal surface. The cell lineage comprises immature pre-pit cells near the proliferative isthmus, mature pit cells within the foveola, and older interfoveolar cells lining the free surface. Ultrastructurally, differentiation can be traced towards a predominantly mucus-producing cell type and finally towards a surface-protective cell variant, which degenerates in situ and is extruded into the lumen without affecting epithelial integrity.

Análisis morfológico comparado de la región fúndica entre un cánido (canis familiaris) y un camélido sudamericano (lama glama) / Comparative morphological analysis of the fundic region between a canidae (canis familiaris) and a southamerican camelidae (lama glama)

Los hábitos alimentarios que presentan los cánidos (monogástricos) respecto de los camélidos (poligástricos), son diferentes. Los primeros consumen dietas pobres en fibras, en tanto que los camélidos consumen una dieta rica en fibra que consiste en pastos cortos y duros (bofedal). Sin embargo son capaces de aprovechar eficientemente estos alimentos. Por ello, fueron analizados comparativamente las regiones fúndicas de ambas especies, donde encontramos diferencias morfológicas adaptativas. Para la realización del presente trabajo, se obtuvieron muestras de las regionnes fúndicas, provenientes de estómagos de perro (Canis familiaris) y de llama (Lama glama), adultos. estas muestras fueron procesadas de acuerdo a las técnicas histológicas corrientes e histoquímica de Picrosirius y analizadas comparativamente al microscopio óptico. Los resultados revelaron que existen diferencias importantes en las regiones fúndicas de ambas especies en estudio. Estas diferencias se encuentran especialmente a nivel de la mucosa, donde la profundidad de las glándulas fúndicas, número de células parietales (oxínticas) y la irrigación sanguínea son mucho mayores en la llama. La pared abdomasal presenta un refuerzo de tejido conjuntivo compacto de haces paralelos, entre la serosa y la muscular longitudinal externa. Estas adaptaciones redundarían en una mayor eficiencia digestiva por parte de los camélidos, dadas las características de su dieta habitual

pH-dependent association of carbonic anhydrase (CA) with gastric light microsomal membranes isolated from bovine abomasum. Partial characterization of membrane-associated activity.

1. The effect of pH on the association of carbonic anhydrase (CA) with bovine gastric light microsomal membranes (LMMs) was investigated (a) by washing LMMs containing CA activity with solutions of different pHs; (b) by studying the adsorption at various pHs of soluble bovine erythrocyte CA to washed gastric LMMs. In both cases, the association of CA with gastric LMMs was dependent on pH, being lower at neutral or alkaline pH. 2. The amount of soluble CA associated with gastric LMMs at pHs 8.0 and 9.0 was reduced when 140 mM K+/10 mM Na+ was added to the incubation medium. 3. Two sources of CA activity in bovine gastric LMMs were assumed: a loosely- and a firmly-membrane-associated activity. Both CA activities were dose-dependently inhibited by acetazolamide (I50: 3.6 x 10(-9) and 8.4 x 10(-9) M, respectively) and by chloride, acetate, iodide, bromide and nitrate at 100 mM. Firmly-membrane-associated activity appeared to be less sensitive to inhibition by acetazolamide, chloride and iodide. 4. Both activities exhibited different behavior and stability following treatment with alkaline Triton X-100. 5. The possible importance of a membrane-associated CA activity in gastric LMMs related to gastric acid secretion is discussed.

Scanning electron microscopy of the mucosal surface of the forestomachs and abomasa of grey, white and black Karakul lambs.

Homozygous grey and white Karakul lambs suffer from a lethal genetic factor causing death after weaning. Previous studies revealed large milk-filled rumens in the grey and white lambs which was attributed to a significant decrease in the number of myenteric ganglia and neurons in the rumen wall. This study was undertaken to determine the effect of milk on the epithelial lining of the forestomachs of affected grey and white lambs. In the forestomachs of the black lambs the polygonal epithelial cells were tightly packed, seemed to overlie one another and cytoplasmic projections occurred on the cell surfaces. In the grey and white lambs the epithelium had an eroded appearance due to sloughing of the surface cells and the cytoplasmic projections were lower and had a weathered appearance compared to the black lambs. No obvious differences could be detected in the abomasa of grey, white and black lambs. It is concluded that the milk in the forestomachs of the grey and white lambs is responsible for the epithelial changes.

Production of prochymosin, pepsinogen and progastricsin, and their cellular and intracellular localization in bovine abomasal mucosa.

A brief overview of research is presented on the production, cellular and intracellular localization of prochymosin, pepsinogen and progastricsin in bovine abomasal mucosa from fetus to adult. Prochymosin is produced early during gestation (10th week) and is significantly related to milk-feeding. Pepsinogen and progastricsin start to be produced later during gestation (20th week) and are produced in low amounts as long as the calf is fed milk. With age, pepsinogen becomes the dominating zymogen in the abomasal mucosa. Most of the cell types in the fundic gland have the ability to produce all three zymogens and are also found in the same individual secretory granules of these cells.

Intracellular enzyme localization in the epithelium of the bovine glandular stomach.

The intracellular localization of calcium adenosine triphosphatase (Ca2(+)-ATPase) was studied ultracytochemically in the pyloric glands of the abomasal mucosa of cattle. A remarkable staining pattern exhibited the Golgi apparatus, as there was a gradation in staining of the interior sides of dictyosomal cisternae from the not or weakly stained cis to the heavily stained trans face. Membranes of Golgi-endoplasmic reticulum lysosome complex-secretory vesicles showed either no or strong enzyme activity. Membranes of secretory vesicles accumulated in the cell apex stained positive for ATPase activity. This accounts also for the apical cortical cytoplasm. From these results it is speculated that Ca2(+)-ATPase may play an important role in the pathway of exocytotic secretion, especially in the process of membrane sorting and biogenesis of secretory vesicles, in the steps of vesicle accumulation and transport to the site of exocytosis as well as in membrane fusion events.

Ultrastructural study of the endocrine cells of the abomasum mucosa in pre-ruminant calves.

During a systematic ultrastructural study on the endocrine/paracrine cells of the gastro-intestinal tract of ruminants, the aim of present work is to describe these cells in the abomasum of suckling calf. Taking into account all the cytological details, and especially the morphological appearance and the cytochemical reactivity of secretory granules, several endocrine cell types can be distinguished. Some of them (EC, D, D1) are common to all three of the glandular regions, whereas others are typical of cardiac and proper gastric glands: they are ECL, X, A-like and a fourth type, whose classification is uncertain. The last type (G cells) is detectable in pyloric glands only. The cardiac and proper gastric glands of the suckling calf abomasum contain two additional cell types, not present in bulls, A-like cells and the fourth type, and contain D1 cells which form a heterogeneous family. These data show a morphofunctional similarity between the abomasum of the suckling calf and the stomach of non-ruminant mammals.

Eight types of endocrine cells in the abomasum of sheep.

In the ovine abomasum, 8 types of endocrine cells were classified at ultrastructural level. The gastric-type EC cells contained oval and pleomorphic granules with high electron density. The intestinal-type EC cells were filled with oval, irregular and highly dense granules. ECL cells contained irregular granules with high density and wide clear spaces. D cells were filled with round granules showing low to moderate density and finely granular matrix D1 cells contained round or oval granules with variable, low to moderate density and finely granular content. G cells showed round and oval granules with moderate density, densely packed or flocculent content. F cells were filled with oval or elliptic granules showing low density with a finely granular and flocculent matrix. X cells contained round granules with high density and homogeneous material. Gastric-type EC cells, intestinal-type EC cells, D cells, and D1 cells were represented in the cardiac, fundic and pyloric gland areas of the ovine abomasum. ECL cells and F cells were confined to the fundic glands, G cells and X cells to the pyloric glands.