Ontogenesis of the rumen: A comparative analysis of the Merino sheep and Iberian red deer

The aim of this study is to describe differences in the ontogenesis of the rumen in the sheep (domestic ruminant) and deer (wild ruminant). A total of 50 embryos and fetuses of Merino sheep and 50 of Iberian deer were used, from the first stages of prenatal life until birth. For the study, the animals were divided into five experimental groups according to the most relevant histological characteristics. The appearance of the rumen from the primitive gastric tube was earlier in the sheep (22% gestation, 33 days) than in the deer (25% gestation, 66 days). In both cases it displayed a primitive epithelium of a stratified, cylindrical, non‐ciliary type. At around 28% gestation in the sheep (42 days) and 26% (67 days) in the deer, the rumen was configured of three clearly‐differentiated layers: internal or mucosal, middle or muscular and external or serosal. In both species the stratification of the epithelial layer was accompanied by modifications in its structure with the appearance of the ruminal pillars and papillae. The pillars appeared before the papillae and the appearance of both structures was always earlier in the deer (pillars: 70 days, 27% gestation; papillae: 97 days, 36% gestation) than in the sheep (pillars: 42 days, 28% gestation; papillae: 57 days, 38% gestation). The outlines of the ruminal papillae appeared as evaginations of the basal zone toward the ruminal lumen, dragging in their formation the basal membrane, the lamina propria and the submucosa. The tegumentary mucosa of the rumen was without secretion capability in the first embryonic phases. From 67 days (26% gestation) the neutral mucopolysaccharides appeared in the deer and at 46 days (30% gestation) in the sheep. In both cases they continued to decrease until birth, this diminution being more pronounced in the deer. Finally, the presence of neuroendocrine and glial cells was detected in the deer at earlier stages than in the sheep.     

Histological development and histochemical localization of enzymes in rumen and reticulum in bovine fetuses

Thirty Holstein fetuses from 100 to 251 d after conception were utilized to study prenatal anatomical development of the epithelium of the rumen and reticulum. Four calves at birth were included in the study for comparison. Tissue sections were frozen and stained to locate specific enzymes. At 100 d, the epithelial layer of the rumen was differentiated into a thin basal zone and a thickened superficial zone of undifferentiated cells. The basement membrane was straight, and in both zones cells were perpendicular to it. At 120 to 141 d, low, primary undulations were detected in the basal zone, basement membrane and underlying lamina propria. At 150 to 166 d, secondary undulations and incipient papillae began to resemble the papillae of mature mucosa. In rumen papillae of 192-d to 215-d fetuses, shallow furrows began to separate papillae apexes from the mass of epithelium. In fetuses 244 to 251 d, the papillae began to be a separate entity. At birth, the basal position of the papillae still remained fused. An incipient separation between the papillae was seen. Several dehydrogenase enzymes, including those associated with the Krebs cycle and reductase associated with energy transformation, were observed in both ruminal and reticular tissue. Alkaline phosphatase activity was localized in the stratum corneum and in blood vessels. Development of the honeycomb configuration of reticular epithelium was evident in the 100-d fetus and progressed rapidly with age.     

Subepithelial elastic fibers in the bovine gastric mucosa

The arrangement of subepithelial elastic fibers in the bovine gastric mucosa was studied by light and electron microscopy, with special reference to the elastic architecture of the cornified papillae. A common feature of their arrangement was that the fibers, derived from the elastic plexus in the deep layer of the lamina propria, extend perpendicularly toward the epithelium giving off the slender branches that end at the epithelial basement membrane. The connecting branches were observed electron microscopically in the form of oxytalan or slender elaunin fibers. The high frequency of fibers was found in the mucosa of the upper part of the ruminal atrium, the ruminal pillars, the reticular cristae and two kinds of cornified unguiculiform and omasal papillae. Each papilla consisted of the core of three-dimensional elastic network and numerous connecting-twigs arising from the superficial points of the network. The mucosae of the ruminal sacs, the floor of the reticular cells and the abomasum except the pyloric part were relatively poor in elastic fibers. These results suggest that the variation in fiber-arrangement may be relevant to the condition of mechanical stress inflicting on the gastric innersurface.     

Characteristics of Filiform, Fungiform and Vallate Papillae and Surface of Interface Epithelium-Connective Tissue of the Maned Sloth Tongue Mucosa (Bradypus torquatus, Iliger, 1811): Light and Scanning Electron Microscopy Study

The study of lingual surfaces and the surface of interface epithelium-connective tissue of the tongue of Bradypus torquatus was performed by employing the light and scanning electron microscopy (SEM) techniques. The results revealed that the rostral part of the tongue presents a round apex and covered by filiform and fungiform lingual papillae and a ventral smooth surface. It was observed that the epithelial layer of the dorsal surface possesses the basal, spinosum, granular and cornified epithelial cells. The lamina propria is characterized by a dense connective tissue forming the long, short and round papillae. Numerous typical filiform papillae are located especially in the rostral part intermingled for few fungiform papillae, which were revealed in three-dimensional SEM images. Usually, the fungiform papillae are located in the border of rostral apex of the tongue exhibiting the rounded form. They are covered by keratinized epithelial cells. In the fungiform papillae, several taste pores were observed on the surface. The vallate papillae presented numerous taste buds in the wall of epithelial cells, being that the major number of taste buds is located on the superior half of vallate papilla. The taste pores are surrounded by several laminae of keratinized epithelial cells. The samples treated with NaOH solution and examined by SEM revealed, after removal of the epithelial layer, the dense connective core in original disposition, presenting different sizes and shapes. The specimens stained with Picrosirius and examined by polarized light microscopy revealed the connective tissue, indicating the collagen fibres type I and type III.     

Histomorphometric analysis of the rumen of sheep during development.

Histomorphometric and scanning electron microscopic analyses were carried out on 74 embryos and fetuses and 20 sheep (early postnatal to adult age). Histodifferentiation of the rumen took place at 33 days of fetal life. Ruminal pillars were observed at 42 days, and at 61 days, ruminal papillae appeared as evaginations of the epithelial stratum basale. Neutral mucopolysaccharides first appeared in epithelial cells at 46 days of fetal life; thereafter, numbers decreased gradually and subsequently stabilized in postnatal life. Acid mucopolysaccharides, mucins, and mucoid compounds were not detected. Age and diet were recognized as factors that determine the structure of the ruminal mucosa. Growth curves and formulas were set out for each tissue layer.     

Histological characteristics of induced acute peptic injury in equine gastric squamous epithelium.

The objective of the study reported here was to characterise the microscopic appearance of peptic-injured equine gastric squamous epithelium in relation to the duration of peptic injury. Erosions and ulcers were induced in equine gastric squamous epithelium using a feed deprivation protocol that results in prolonged increased gastric acidity. Specimens of normal gastric mucosa and mucosa with lesions created after 48 and 96 h of feed deprivation were compared for characteristics associated with angiogenesis and mucosal proliferation. Fifteen mature horses, 9 geldings and 6 mares, age 3-20 years, were divided into 3 groups. Group 1 (n = 5) had normal-appearing gastric squamous mucosal epithelium and had been killed due to problems unrelated to the gastrointestinal tract. Groups 2 (n = 5) and 3 (n = 5) had lesions induced in the gastric squamous epithelium by alternating 24 h periods of feed deprivation and ad libitum access to hay, for totals of 48 and 96 h feed deprivation, respectively. Following lethal injection of barbiturate, stomachs were removed and fixed by filling with 4-6 l 10% buffered formalin. Sections were made from lesions in the gastric squamous epithelium adjacent to the margo plicatus along the right side of the stomach/greater curvature and the lesser curvature. Measurements of total epithelial thickness, keratinised epithelium, nonkeratinised epithelium, epithelial projections, capillary extension into the epithelium and lamina propria thickness were made. The cross-sectional areas of arterial and venous vascular structures in the lamina propria at the lesions and their margins were measured using image analysis software. All horses, except one, in Group 2 developed erosions or ulcers in the gastric squamous epithelium after feed deprivation. There were several changes in the epithelium adjacent to erosions and ulcers, compared to normal epithelium, from horses in Groups 2 and 3: total epithelial thickness was significantly (P<0.05) greater, including both keratinised and nonkeratinised layers in most specimens; the length of epithelial projections and extent to which capillaries from the lamina propria extended toward the luminal surface, and the cross-sectional area of vascular structures (arterioles, capillaries, venules) in the lamina propria were significantly greater. Epithelial thickness of erosion beds was not significantly less than normal epithelium, although a greater proportion of the epithelium in erosions consisted of epithelial projections (Group 1, 23%; Group 2, 76%; Group 3, 72%). The cross-sectional area of vascular structures in the lamina propria beneath erosions was significantly greater than in normal mucosa only in Group 2 tissues, whereas in the lamina propria of ulcers it was significantly greater than in normal mucosa only in Group 3 tissues. The epithelial proliferation and increased vascular cross-sectional area in the lamina propria associated with peptic-induced gastric lesions are consistent with processes associated with the initiation of ulcer healing, and these changes temporally coincided with the initiation of peptic insult to the gastric squamous epithelium. These findings demonstrate that processes that promote ulcer healing begin soon after peptic injury and that they progress even with repeated peptic injury. Furthermore, our findings support observations that gastric ulcers often heal without medical intervention, and the theory that medications that reduce gastric acidity do not initiate healing, but rather facilitate ulcer healing by providing a microenvironment that is optimal for healing to proceed.     

Is Lamina Muscularis Mucosae Present in the Ruminal Mucosa of Cattle?

The cells in the condensed fibrous layer in the lamina propria of the ruminal mucosa of the calves and cows were studied at the light microscopic level. Abundant actin-immunoreactive cells were detected both in the calves and cows, while desmin-immunoreactive cells were moderate in frequency in the cows and low or absent in the calves. Vimentin-immunoreactive cells were abundant in the cows but moderate to low in frequency in the calves. These cells were distributed to form a condensed layer being more abundant in the ruminal papillae than the interpapillar mucosae in the cows and equal in the papillae and interpapillar mucosae in the calves. At the ultrastructural level, smooth muscle cells and fibroblasts were abundant in the deep core of the ruminal papilla of the cows. The present results suggest that the condensed fibrous layer revealed in the ruminal mucosae contains smooth muscle cell population which could be regarded as the specialized lamina muscularis mucosae.     

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.     

Einfluß der Ernährung auf den Papillarkörper der Pansenschleimhaut bei Kälbern

The purpose of this scanning electronmicroscopical study was to describe the structure of the lamina propria of the ruminal mucosa in calves on different diets.
The surface epithelium was removed with 0.3 % acetic acid. The exposed lamina propria showed structural differences, both macroscopically and microscopically in the different dietary groups. At the submicroscopic level, however, no differences were observed.     

Preliminary observations of interaction between bacteriophages and Streptococcus bovis bacteria on ruminal epithelium primoculture.

Five Streptococcus bovis strains (47/3, 59/2, 4/1, 46/2 and 44/9) isolated from calf ruminal fluid samples were examined for the adherence to cultured ruminal epithelium cells. Four strains (47/3, 59/2, 4/1 and 46/2) were able to attach to the cultured epithelial cells. However, S. bovis 47/3 strain attached to the target cells in significantly greater numbers than the other strains. Strain 44/9 did not adhere to cells of ruminal epithelium. The adherent bacteria were observed on the surface of differentiated (mainly keratinized) cells of ruminal epithelium primoculture only. The different effect of F4, F5 and F6 bacteriophages was ascertained on S. bovis bacteria adhering to rumen epithelial primoculture. A significant decrease in the number of adherent bacteria was shown after cultivation of strains 47/3 and 4/1 with F6 bacteriophage and of 47/3 strain with F4 phage. The F5 bacteriophage had no significant effect on these bacteria.     

Morphological study of the Forestomach of the Japanese Serow (Capricomis crispus)

Details of the morphology of the forestomach of the Japanese serow (Capricornis crispus) were examined macroscopically, and by light and scanning electron microscopy. The rumen was sac-like in shape, and the right accessory longitudinal groove was not visible externally. The reticulum was relatively small, while the omasum was oval and large. Inside the rumen, even in the dorsal sac, the ruminal papillae were densely distributed, although absent on the pillars. The caudodorsal coronary pillar was indistinct. In the ruminoreticular fold, ruminal papillae were observed even on the reticulum side. In the reticulum, the primary reticular fold was apparent and the ill-defined secondary folds were also detected in some reticular cells. In the omasum, the omasal laminae of four sizes were recognized. The fourth order of laminae was occasionally absent. Histological sections of ruminal papillae revealed that softly keratinized epithelium covered the entire ruminal mucosa. In the reticulum, soft keratinized epithelium was observed throughout, with the exception of the tips of reticular papillae on which the epithelium showed evidence of hard keratinization. The epithelium of the omasal laminae was moderately keratinized. On the floor of the reticular groove, interlaminar area and in the omasal groove, the accumulations of numerous eosinophilic cells were observed in the superficial region. Under the scanning electron microscope, shallow transverse grooves were seen on the surface of the ruminal papillae. In general, the surface structure seemed to be similar to that of sheep rather than cattle. Cellular formations of the collagen fibrils were observed at the subepithelial region. In conclusion, examination of the forestomach allows classification of the Japanese serows as requiring an intermediate type of feeding habit and being between concentrate selectors and grass-roughage eaters from a morphological view.     

Development of the Bovine Duodenum with Reference to Enterochromaffin cells

The present light microscopic study describes the morphological changes which occur in the duodenal mucosa of the bovine fetus from the third month of gestation until birth with regard to the sequence of appearance and morphology of enterochromaffin cells. The mucosa of the duodenum is prepared for digestive activity during the first months of development. This becomes obvious in the early differentiation of the fetal intestinal mucosa. The enterocytes in villus epithelium contain glycogen in the early months. These glycogen pools seem to shrink during the following months. They may provide a metabolic reserve for the fetus, which derives all of its nutrition from carbohydrates. Enterochromaffin cells appear in the three month old bovine fetus with two cell-types situated in the epithelial lining of crypts and villi. The density of enterochromaffin cells increases during gestation. There were no findings to indicate that the fetal enterochromaffin cells did not originate from endoderm. The early appearance from enterochromaffin cells (which are the producers of serotonin, 5-hydroxytryptamine) correlates with first gastrointestinal movements, which are known to commence in the first third of gestation, leading us to believe that 5-HT represents the chemical inducer of muscular motility in the developing intestine.     

Histomorphometric study of the goat stomach during prenatal development

The aim of this study was to perform a morphometric analysis of the different structural tissue layers of the goat stomach to study their prenatal growth from mathematical models fitted to these morphometric data. A total of 90 embryos and fetuses were used, from the early stages of prenatal life until birth. The growth rate of the gastric wall was slower than that of body length; rumen was the stomach compartment displaying slowest growth. In the three non‐glandular compartments, the epithelial layer grew faster than the gastric wall itself, while the growth rate of the abomasal epithelium declined in the early stages of development. A decline in growth rate was also observed for the lamina propria and submucosa in rumen and reticulum from the early embryonic stages, whereas in omasum and abomasum these layers continued to grow as gestation progressed. The tunica muscularis displayed consistent growth in all compartments, growing faster than the gastric wall. Serosa thickness increased as gestation progressed, displaying a decline in growth‐rate only in the omasum. In conclusion, the dynamics of gastric wall growth were governed by the growth rate of each of the component tissue layers.     

山羊咽扁桃体和咽鼓管扁桃体的组织结构观察

选取健康10月龄奶山羊10头,断头宰杀后取咽扁桃体和咽鼓管扁桃体,应用组织学光镜和电镜制片技术研究咽扁桃体和咽鼓管扁桃体的显微和亚显微组织结构.结果表明:山羊咽扁桃体和咽鼓管扁桃体的黏膜上皮主要由2～3层多边形上皮细胞组成,部分区域只有单层扁平细胞,相邻上皮细胞间空隙很大,上皮细胞表面有丰富的微绒毛.上皮细胞之间和黏膜上皮下方固有层内有大量淋巴细胞浸润.扁桃体的实质部分由数个次级淋巴小结和弥散淋巴组织构成,弥散淋巴组织中有大量分布的淋巴管和毛细血管后微静脉.此外,在紧贴黏膜上皮细胞下方的固有层和淋巴滤泡中可观察到少量的树突状细胞.结果提示山羊的咽扁桃体和咽鼓管扁桃体可作为鼻腔免疫的主要诱导位点和效应部位.     

Fine structure of developing gastric parietal cells in the bovine abomasum

Developmental changes in the fine structure of gastric parietal cells were studied in bovine fetuses and neonates. Definitive parietal cells first appeared as non-granular light cells at five months of fetal age (in a 48 cm long fetus). At this age they had a microvillous apical surface and cytoplasmic vesicles which accumulated at the apex. Intracellular canaliculi appeared with an increase in mitochondria at six to seven months of fetal age. Fully differentiated parietal cells were present from the eighth month of gestation. At birth they were similar in appearance to those of the adult abomasum.     

Factors influencing the adherence of strains of Streptococcus bovis and Escherichia coli isolated from ruminal epithelium

Two strains of Streptococcus bovis (A1 and A5) and one strain of Escherichia coli (0141: H28) isolated from the surface of bovine ruminal mucous epithelium were examined for adherence to isolated and cultured ruminal epithelial cells. The E. coli adhered to the target cell by means of fimbriae, which had several common properties with type 1 common fimbriae and caused mannose-sensitive haemagglutination. The A1 strain of S. bovis was devoid of fimbriae and its adherence to the epithelial surface was not inhibited by treatment with sugars or phenol-treated bacterial membrane from the same organism. It was therefore postulated that the bacterial glycocalyx of the S. bovis organisms acted as ligand. The extent of bacterial adherence depended on the state of differentiation of the target cell in both the isolated and the cultured ruminal cell systems. The receptors for both adherent bacterial species were in all probability associated with the glycocalyx of the target cells.     

Mucin biosynthesis in the bovine goblet cell induced by Cooperia oncophora infection

Mucin hypersecretion is considered to be one of the most common components of the immune response to gastrointestinal nematode infection. However, investigations have not been conducted in the Cattle–Cooperia oncophora system to verify the findings largely derived from murine models. In this study, we examined the expression of seven mucins and seven enzymes in the mucin biosynthesis pathway involved in O-linked glycosylation in the bovine small intestine including goblet cells enriched using laser capture microdissection during a primary C. oncophora infection. At the mRNA level, MUC2 expression was significantly higher in both lamina propria and goblet cells at 28 days post-infection compared to the naïve control. MUC5B expression at the mRNA level was also higher in lamina propria at 28 dpi. Expression of MUC1, MUC4, MUC5AC, and MUC6 was extremely low or not detectable in goblet cells, columnar epithelial cells, and lamina propria from both naïve control and infected animals. Among the seven enzymes involved in post-translational O-linked glycosylation of mucins, GCNT3, which may represent one of the key rate-limiting steps in mucin biosynthesis, was up-regulated in goblet cells, columnar epithelial cells, lamina propria, and gross small intestine tissue during the course of infection. Western blot analysis revealed that MUC2 glycoprotein was strongly induced by infection in both gross small intestine tissue and its mucosal layer. In contrast, the higher MUC5B protein expression was observed only in the mucosal layer. Immunohistochemistry provided further evidence of the mucin glycoprotein production and localization. Our results provided insight into regulation of mucin biosynthesis in various cell types in the bovine small intestine during gastrointestinal nematode infection and will facilitate our understanding of mucins and their role in immune response against parasitic nematodes.     

Morphological Studies of the Tongue of the Egyptian Water Buffalo (Bubalus bubalis) and Their Lingual Papillae Adaptation for Its Feeding Habits

This work was conducted to describe the morphological characters of the tongue of Egyptian water buffalo (Bubalus bubalis). The lingual root and the dorsal middle region of apex and body in addition to the dorsal and ventral surface of lingual tip were devoided from any fungiform papillae. The lingual tip contains conical papillae only. The ventral surface of lingual apex was divided into two portions by the U‐shaped fungiform line into papillary and non‐papillary region. Histological investigation on the lingual surface epithelium and lamina propria submucosa reflects differences in these layers in different parts of the tongue. By SEM, there are two subtypes of filiform papillae: caudally directed papillae on dorsal surface and rostrally directed papillae on the lateral region of ventral surface of lingual apex. There are two subtypes of conical papillae: small slightly rostrally directed papillae on dorsal and ventral surface of lingual tip and large posteromedially directed papillae on dorsal surface of lingual root. The rounded circumvallate papillae consisted of round bulb surrounded by deep circular groove, which surrounded by circular pad. Higher magnification of filiform papillae indicates the presence of microcrests separated by microgrooves, and these microgrooves consisted of microrodes. The fungiform surface having micropores on the tip of elevated tubercle for taste buds pores. All these observed structures (microcrests, microgrooves, microrodes, tubercles, microridges) in a higher magnification allow animals to transport food particles through the oral cavity and help in the defensive behaviour. There are strong correlations between the tongue anatomical characteristics and its functions.     

Histomorphological features of the tongue of the Eurasian teal (Anas crecca)

This study presents the histomorphological features of tongue in Eurasian teal (Anas crecca); the smallest extant dabbling duck. Heads of four adult males and four adult females were used in this study. The results illustrate a tongue with three different parts; the apex with a lingual nail in ventral surface, the body with a lingual prominence in caudal part and some large and small conical papillae in lateral sides and the root, that was covered with many conical papillae in different sizes. Histological results revealed two types of keratinized and non‐keratinized epithelium covering parts of the tongue. The lingual salivary glands were observed in the lamina propria of the body and root of the tongue showed strongly periodic acid–Schiff (PAS)‐positive reaction. The yellow adipose tissue was located under the lamina propria on the body and root of the tongue. The filiform papillae between the conical papillae of the body were arranged densely. The sensory organs, which contain sensory receptors (Grandry and Herbst corpuscles), were located in the lamina propria of the body of the tongue. In conclusion, the anatomical and histological structure of the Eurasian teal’ tongue was generally similar to its family members such as domestic goose and duck but showed some differences that may be adoptions to the bird's habitat and mode of feeding.     

Morphological and functional development of the rumen in the calf: influence of the time of weaning. 1. Morphological development of rumen mucosa.

The objective of this study was to determine whether the nutritional regimen of rearing calves would influence the morphometric and histological development of rumen mucosa. Twelve male Holstein calves 7 d of age were assigned to three groups of 4 animals each: milk group (I), early weaned (6 weeks) group (II) and late weaned (9 weeks) group (III). All animals received additional solid feed. Animals of group I were slaughtered after 6 weeks of age, whereas those in groups II and III were slaughtered after 9 weeks of age. At slaughter, the ruminal digesta amounted to 2035 g (milk group), 3092 g (late weaned group) and 5374 g (early weaned group). The differences in the ruminal molar percentage of SCFA were not significant. There was a trend for lower pH and higher SCFA concentrations in the order late weaned, early weaned and milk fed animals (pH: 6.4, 6.6 and 6.7, respectively; SCFA: 96, 87 and 77 mmol/l, respectively). The mean length (1.07 mm in milk group, 1.45 mm in late weaned group and 1.87 mm in early weaned group), width (0.43, 0.58 and 0.71 mm, respectively) and surface of papillae (190, 232 and 241 mm2/cm2 mucosa, respectively) increased with both the age of the animals and the elevated intake of solid feed, whereas the number of papillae (210, 140 and 92 per cm2 mucosa, respectively) decreased. In both milk-fed groups type A and B corneal cells were present in the Stratum corneum, whereas in the earlier weaned calves type C-cells could be also seen. These findings indicate a more advanced stage of development of the rumen epithelium in the earlier weaned calves fed higher amounts of concentrate and hay.