Lectin histochemistry of trachea and lung of healthy turkeys (Meleagris gallopavo) and turkeys with pneumonia

Thirteen lectins were used to characterize lectin-binding specificity of glycoconjugates on sections of formalin-fixed lung and trachea from seven normal turkeys, two turkeys with acute pneumonia, and two turkeys with chronic pneumonia. Neuraminidase was used to digest sialic acid residues. One N-acetylgalactosamine-binding lectin and two N-acetylgalactosamine/galactose-binding lectins stained the apical membrane and cytoplasm of multifocal cells that lined air atria and hyperplastic granular cells. Other lectins in these groups stained ciliated cells of the trachea and bronchi and air capillary epithelial cells. Sialic acid residues were on apical surfaces of ciliated and nonciliated tracheal and bronchial lining cells, air capillary epithelial cells, and vascular endothelial cells. Mannose/glucose-binding lectins stained reticular and elastic fibers in the lamina propria of trachea, primary and secondary bronchi, and the tunica adventitia of arteries and veins. By transmission electron microscopy, colloidal gold-Arachis hypogaea (peanut agglutinin) labeled microvilli on the apical surface of mature granular cells. The L-fucose-binding lectin, in addition to several other lectins, stained nonspecifically in both trachea and lung. These studies show that granular cells that line air atria can be identified with lectins of N-acetylglucosamine and N-acetylgalactosamine/galactose groups, and that apical surfaces of epithelial cells and endothelial cells in the trachea and lung express terminal sialic acid residues.     

Plasma 1,25 dihydroxycholecalciferol and its free index are potentiated by ovulation dependent factors and shell formation induced hypocalcemia in the laying hens.

The time-course of the changes in blood ionized calcium, and in plasma 1,25 dihydroxycholecalciferol (1,25(OH)2D3) concentrations and its free index were studied in hens following suppression and resumption of shell formation and throughout the laying cycle in hens laying hard-shelled eggs, in hens fed a low or normal calcium diet and in hens laying shell-less eggs. The respective roles of the calcium needs for shell formation and of the reproductive status in regulation of 1,25(OH)2D3 production were analysed. Plasma 1,25(OH)2D3 decreased 3 hr after suppression of shell formation following premature egg expulsion and remained lower than that of hens laying hard-shelled eggs when premature expulsion of the eggs was continued for several days. Circulating 1,25(OH)2D3 tended to increase progressively when shell formation was resumed. Ablation of the parathyroid glands abolished this increase. In hens laying hard-shelled eggs, the plasma 1,25(OH)2D3 was higher during the period of shell secretion. Feeding hens a low calcium diet (1.2%) caused a marked increase in the plasma 1,25(OH)2D3. Ionized calcium levels tended to show reciprocal changes to plasma 1,25(OH)2D3 decreasing when calcification took place and increasing after its suppression. In hypercalcemic hens laying shell-less eggs and fed a 3.5% Ca diet, the plasma 1,25(OH)2D3 was at a high level 4 hr after ovulation and diminished thereafter. This additive stimulation does not, therefore, involve the parathyroid gland and may involve hormonal changes induced by ovulation. Vitamin D binding protein (DBP) in the plasma was at a high level in mature hens and was not affected by shell formation. Consequently, the free 1,25(OH)2D3 index fluctuated in parallel with total level of this hormone in mature hens. It is concluded that the calcium demand for shell formation modulates, in the short term, plasma 1,25(OH)2D3, via the homeostatic regulation of blood calcium by PTH, but that a large part of its increase is independent of PTH and is associated with the endocrine events concomitant with ovulation.     

Microscopic anatomy of the lower respiratory tract of the grey short-tailed opossum (Monodelphis domestica)

The respiratory tracts of seven grey short-tailed opossums were histologically examined. Six opossums were prepared by perfusion with buffered formalin. Opossum seven was perfused with gluteraldehyde. Samples taken from the respiratory passages and lungs of specimens 1-6 were stained with haematoxylin and eosin. A mixture of methylene and azure blue was used for specimen 7. The trachea and right and left principal bronchi are lined with a pseudostratified ciliated columnar epithelium with occasional goblet cells. The secondary and tertiary bronchi and the primary and secondary bronchioles are lined by a simple ciliated columnar epithelium. The terminal bronchioles and a portion of the respiratory bronchioles are lined by a simple ciliated cuboidal epithelium. The terminal portion of the respiratory bronchioles and the alveolar ducts are lined with simple squamous epithelium. Alveoli are lined by type I and II pneumocytes. Tracheal glands are present in the tela submucosa. The fibromusculocartilaginous tunic of the trachea consists of c-shaped cartilage rings and the trachealis muscle. A lamina muscularis mucosa begins in the intrapulmonary portion of the principal bronchus and continues into the respiratory bronchioles. Bronchial glands are present in the propria submucosa and tela submucosa of the principal bronchi. The musculocartilaginous tunic is localized to the extrapulmonary portion of the principal bronchus. The bronchial cartilages are irregular shaped plates and limited to the extrapulmonary portion of the principal bronchus. The visceral pleura is a simple squamous mesothelium covering the outer surface of the lung.     

Superficial eosinophilic gastritis in laboratory beagle dogs attributable probably to diet

Stomach tissues from 222 beagle dogs, which were used for toxicological studies, were examined histopathologically. Out of 190 dogs fed the same diet, 185 had gastric lesions, whereas all of 32 dogs fed another diet were free from them. The gastric lesions were noted as an accumulation of eosinophils, infiltration of inflammatory cells including lymphocytes, plasma cells and histiocytes in the gastric lamina propria, formation of foreign body type multinucleated giant cells, focal desquamation and papillary proliferation of mucosal epithelium. The lesions were confined to the superficial layer of the antral and pyloric mucosa, and neither desmoplasia nor clinical signs were seen. Multinucleated giant cells often contained amphophilic foreign bodies of varying appearance with inconsistent reaction for periodic acid-Schiff (PAS), but no definite causal agents were detected.     

Giant cell tumour of bone in a cat with extraskeletal metastases: pathological and immunohistochemical study

A case of giant cell tumour of bone (GCTb) in the lung and in a subcutaneous mass located in the right flank, with a probable primary origin in the mid-diaphysis of the right tibia, was described in a 8-year-old female cat. Numerous multinucleated giant cells were homogeneously distributed among a population of ovoid or spindle-shaped mononuclear cells. All of them were positive for vimentin suggesting a mesenchymal origin. Spindle-shaped tumour cells resemble fibroblastic cells, showing collagen fibres in their vicinity. Ovoid mononuclear cells are similar to macrophages, with a cytoplasm rich in electron-dense lysosomes. Multinucleated giant cells appear morphologically similar to osteoclasts. These findings are supported for the positive reaction to tartrate-resistant acid phosphatase (TRAP) and lysozyme, encountered only in ovoid and multinucleated giant cells. No immunoreactivity against human oestrogen receptors was observed in the nuclei of any neoplastic cells.     

Osteoclast progenitors from cats with and without tooth resorption respond differently to 1,25-dihydroxyvitamin D and interleukin-6

Both vitamin D and inflammatory cytokines can stimulate osteoclast formation and activity. We studied the effect of 1,25-dihydroxycholecalciferol (1,25(OH)(2)D), and interleukin-6 (IL-6), on the formation and activity of feline osteoclasts, using peripheral blood mononuclear cells (PBMCs) from cats with and without tooth resorption (TR(+) and TR(-)) as a source of osteoclast precursors. The formation of osteoclast-like cells (defined as multinucleated, tartrate-resistant acid phosphatase-positive cells) was assessed at 7 and 14 days. In the presence of M-CSF and RANKL, with and without IL-6, more osteoclasts were formed from TR(-) PBMCs than from TR(+) PBMCs on plastic. More osteoclasts were formed from TR(+) PBMCs on bone slices in the presence of M-CSF/RANKL with 1,25(OH)(2)D. This opposite effect may be due to a higher expression of the vitamin D receptor (VDR) in TR(+) osteoclasts and precursors on bone. Formation of resorption pits was analyzed and confirmed with scanning electron microscopy. In conclusion, we propose that TR(+) PBMCs when cultured on bone are sensitive to 1,25(OH)(2)D, whereas the differentiation of TR(-) PMBCs on bone seem more sensitive to IL-6, suggesting that osteoclast precursors from cats with and without tooth resorption respond differently to osteoclast stimulating factors.     

Effect of 1,25 dihydroxyvitamin D3 on the calcium and magnesium metabolism of lactating cows

Administration of 1,25 dihydroxyvitamin D3 (1,25(OH)2D3) to lactating dairy cows resulted in increased dietary calcium absorption and elevated concentrations of plasma calcium. Dietary magnesium absorption was unaffected by 1,25(OH)2D3 however, plasma magnesium concentration was depressed. Injections of 1,25(OH)2D3 were effective in elevating plasma calcium concentrations in both normal and hypomagnesaemic cows. This indicates a potential use for 1,25(OH)2D3 to prevent and treat hypocalcaemic cows with or without concurrent hypomagnesaemia.     

Light and electron microscopic studies of the trachea in the one-humped camel (Camelus dromedarius)

Histology of trachea of camel (Camelus dromedarius) was studied using light, scanning (SEM) and transmission electron microscopy (TEM). Tissue samples taken from the trachea (proximal, middle and distal part) were routinely prepared for histology (LM, EM) and stained with haematoxylin and eosin, Van Giesson (VG), Alcian blue, Periodic acid schiff (PAS), Masson's trichrome (MT), Verhof, PAS-VG and PAS-MT. The trachea of camel consists of 66-75 incomplete cartilaginous rings of hyaline. The lamina epithelium is composed of pseudostratified-ciliated columnar epithelium with many goblet cells. Submucosal layers were loose connective tissue with many elastic fibres. The mucosal and submucosal layers were 517.2 +/- 61.6 microm (n = 20) thick. Submucosal glands were tubuloalveolar with mucous (acidic and neutral) secretions. Trachealis muscle was attached to the inside sheet of tracheal cartilage. Ultrastructural studies showed that surface epithelium is pseudostratified with mucus-producing goblet cells, ciliated and basal cells, similar to other mammals. The ciliated cells contained many mitochondria, oval nucleus and many big granules. In scanning electron microscopy (SEM) studies, viscoelastic layers were observed on the epithelial surface of trachea, and there were highly condensed cilia under this layer.     

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.     

雌性空怀双峰驼生殖道的形态结构

应用组织学方法观察了雌性空怀双峰驼生殖道的形态结构。结果显示，双峰驼生殖道的基本结构与其他哺乳动物相似，但微细结构有差异。双峰驼输卵管粘膜皱襞极其发达，分支多而呈复杂的网状迷路。皱襞基部的迷路酷似固有膜而存在腺体，迷路网格内常见细胞团块。虽然双峰驼怀孕时胎儿位于左侧子宫角，但左、右子宫角以及子宫体的组织结构基本相同。子宫内膜无肉阜，上皮下陷于固有膜内，形成大量长而弯曲的单管状腺。子宫颈固有膜浅层分布有许多小腺体，深层分布有成群的较大腺体。这些腺体为分支管状腺，腺上皮PAS强阳性。阴道粘膜上皮为复层上皮。从输卵管到阴道，粘膜上皮主要为单层柱状上皮，由纤毛细胞和分泌细胞组成，局部可见假复层柱状纤毛上皮。纤毛细胞由前向后逐渐减少，但在子宫颈仍可见到。粘膜上皮和腺上皮内夹有许多淋巴细胞或中性粒细胞，后段局部甚至见到这些免疫细胞浸润于上皮细胞间。固有膜内分布有大量淋巴细胞、中性粒细胞、肥大细胞、浆细胞和巨噬细胞，有时出现淋巴滤泡。     

犬正常发情期与患子宫蓄脓的子宫和卵巢组织结构及乳铁蛋白表达的对比

本研究旨在探明犬子宫和卵巢在正常发情期与患子宫蓄脓时，其组织结构及乳铁蛋白（lactoferrin，LF）表达的变化特征。应用Masson’s、VVG、PAS组织化学染色方法观察乏情期、发情期、患子宫蓄脓时犬子宫及卵巢的组织结构特点，用免疫组织化学SP法观察LF的分布特征。结果显示，正常发情期犬：乏情期、发情期子宫内膜肌层厚度比分别为0.762 0、0.924 3；乏情期子宫固有层中胶原纤维含量大于发情期，发情期卵巢中胶原纤维含量大于乏情期；子宫血管层及卵巢血管内弹性膜清晰完整；子宫腺在乏情期时浅层管腔小，深层较大、腺管上皮为单层柱状上皮、上皮细胞及管腔内PAS阳性反应较强，发情期子宫腺管腔变大、腺管上皮为单层立方上皮、上皮细胞及管腔内有PAS阳性反应；子宫黏膜上皮在乏情期和发情期均为单层柱状上皮，但胞核位置不同，乏情期胞核位于中央，发情期胞核位于顶部。患子宫蓄脓犬：子宫内膜肌层厚度比为1.615 0；子宫固有层和卵巢中胶原纤维含量少于正常发情期；子宫腺管腔大，且形状不规则，管腔内有炎性细胞浸润，腺管上皮为单层立方上皮，有淋巴细胞位于基膜，上皮细胞及管腔内PAS阳性反应较弱；子宫血管层及卵巢血管内弹性膜较正常发情期变薄，且有断裂现象；子宫黏膜上皮为单层柱状上皮，胞核位于基底。LF在乏情期子宫腺上皮和卵巢中的表达水平高于发情期，而在子宫黏膜上皮中发情期的表达水平高于乏情期。患子宫蓄脓时犬子宫和卵巢中LF的表达水平均较低。综上表明，犬正常发情期与患子宫蓄脓时的子宫和卵巢组织结构特点显著不同，LF的表达水平也存在差异。     

Scanning electron microscopic study of the lower respiratory tract in calves and adult cattle

The surface characteristics of the lower respiratory tract of two groups of cattle were studied with the scanning electron microscope. Group A comprised six one-week-old calves and group B four adult cows. None of the animals had overt respiratory disease or gross morphological evidence of pulmonary lesions. The trachea, bronchi, bronchioles and alveoli of the cranial and the caudal lobes of the right lung were examined. In both groups the luminal surface of the trachea and large bronchi were completely covered by cilia, apparently forming an efficient mucociliary escalator. In the adult animals there were some patchy areas in the trachea and large bronchi devoid of ciliated cells; these were considered abnormal. In the bronchi, non-ciliated cells, mainly mucus-secreting, were not easily identified unless they were discharging secretion. In small bronchi, non-ciliated cells were more evident and easily seen. The bronchioles had many non-ciliated cells and very few ciliated cells capable of forming a complete carpet for a mucociliary escalator. Types 1 and 2 alveolar epithelial cells and alveolar macrophages were identified in both groups. Pores of Kohn were found in the alveolar walls in all animals.     

A morphologic and immunohistochemical study of the bronchus-associated lymphoid tissue of pigs naturally infected with Mycoplasma hyopneumoniae

Porcine enzootic pneumonia (PEN), caused by Mycoplasma hyopneumoniae (Mh), has been described in pigs in all geographic areas. The disease is characterized by high morbidity and low mortality rates in intensive swine production systems. A morphologic and immunohistochemical study was done to determine the cellular populations present in lung parenchyma of infected pigs, with special attention to the bronchus-associated lymphoid tissue (BALT). Polyclonal and monoclonal antibodies were used for the detection of antigens of Mh, T lymphocytes (CD3+, CD4+, and CD8+), IgG+ or IgA+ lymphocytes, and cells containing lysozyme, S-100 protein, major histocompatibility complex class II antigen or myeloid-histiocyte antigen. Findings in lung tissues associated with Mh infection were catarrhal bronchointerstitial pneumonia, with infiltration of inflammatory cells in the lamina propria of bronchi and bronchioles and alveolar septa. Hyperplasia of mononuclear cells in the BALT areas was the most significant histologic change. The BALT showed a high morphologic and cellular organization. Macrophages and B lymphocytes were the main cellular components of germinal centers. T lymphocytes were primarily located in perifollicular areas of the BALT, lamina propria and within the airway epithelium, and plasma cells containing IgG or IgA at the periphery of the BALT, in the lamina propria of bronchi and bronchioles, in alveolar septa, and around bronchial submucosal glands. The hyperplastic BALT in PEN cases consisted of macrophages, dendritic cells, T and B lymphocytes, and IgG+ and IgA+ plasma cells. CD4+ cells predominated over CD8+ cells. Local humoral immunity appears to play an important role in the infection.     

Microscopic anatomy of the ringed seal (Phoca hispida) lower respiratory tract

The microscopic anatomy of the ringed seal lung exhibits unique features and many features similar to those described in other seal species. Unique features include: Trachealis muscle predominately oriented longitudinally; Large veins within the tracheal wall supported by elastic fibers; Goblet cells and pseudostratified epithelium lining the duct system of bronchial glands of the segmental bronchi; Lamina propria of the segmental bronchus heavily invested with elastic fibers clustered into dense longitudinal bundles; and Capillaries and venules covered with squamous epithelium protruding into bronchiolar lumina. Common features include: Cartilage support of the bronchial tree extending distally into respiratory bronchioles; Smooth muscle enhancements in the distal airways producing sphincter like formations; and Lungs extensively supported with interstitial tissue, which divide lungs into lobules.     

Mast cells in the bovine lower respiratory tract: morphology, density and distribution

The lower respiratory tract of six 8-day-old calves and six adult cows was fixed in either isotonic formal-acetic-acid or neutral buffered formalin in order to study the morphology, density and distribution of mast cells. They were found at all levels of the tract with the highest density in the major bronchi. Tissues from cows had significantly more mast cells than those from calves. There were significantly more mast cells detected in calf airways fixed with isotonic formal-acetic-acid compared with those seen in the same tissues fixed with neutral buffered formalin. Regardless of the age and fixation, mast cells were located predominantly in the alveolar septa and in the lamina propria of airways. They were also commonly encountered within the mucosal epithelium of the trachea. Ultrastructurally, mast cells in bovine airways and lung contained two types of intracytoplasmic granules as described in other species.     

Electron microscopic observation of the respiratory tract of SPF piglets inoculated with Mycoplasma hyopneumoniae.

Seven hysterectomy derived piglets were repeatedly challenged with Mycoplasma hyoneumoniae during the first week of life. Samples of trachea, bronchi and lung tissue collected 2–11 weeks post-inoculation (p.i.) were examined using light and electron microscopy. Autoradiography was used to study in more detail the site of M. hyopneumoniae multiplication. Gross lesions were observed in lung tissue and were characterized by hyperplasia of the epithelium and an increased mononuclear cell accumulation in perivascular and peribronchiolar areas. Mild lesions of the trachea and the bronchi, including epithelial hyperplasia and infiltration of the lamina propria by inflammatory cells, were noted.

Electron microscopy showed that, 2–6 weeks p.i., changes in the mid-trachea and bronchi surface consisted of the loss of cilia. Mycoplasmas covered tufts of cilia remaining on the epithelial cell surface. Scanning and transmission electron micrographs showed that they were predominantly found closely associated with the top of cilia. No specialized terminal structure could be seen and no mycoplasma cells were identified lying free in the lumen nor in close contact with the plasma membrane of cells of microvilli. Some fine fibrils radiating from one mycoplasma to another or to cilia were seen at higher magnification by scanning electron microscopy. Six to eleven weeks p.i., a disrupted epithelial surface lacking cilia was observed. Cells were desquamated and shed into the lumen with cellular remains containing droplets of mucus.

Autoradiography revealed that label corresponded to the observed mycoplasma distribution. At the top of cilia, a high density of labeling was visible in the zone of high mycoplasma concentration. Therefore, incorporation of the label in the mycoplasma is proof or their multiplication in the trachea.

The intimate association between the mycoplasma and cilia may be an important factor in the pathogenesis of the disease caused by M. hyopneumoniae (swine enzootic pneumonia).     

Pulmonary cryptosporidiosis in simian immunodeficiency virus-infected rhesus macaques

Cryptosporidiosis is a common opportunistic infection in the gastrointestinal tract of human and nonhuman primates with AIDS. Pulmonary infection associated with Cryptosporidium spp. has not been previously reported in monkeys. Two macaques experimentally infected with simian immunodeficiency virus (SIV) had lesions containing cryptosporidial organisms involving the trachea, lungs, bile ducts, pancreas, and intestine. The pulmonary sections revealed moderate to severe bronchopneumonia associated with cryptosporidiosis. Numerous 2-4 microm oval Cryptosporidium spp. organisms were present in the cytoplasm of alveolar macrophages, multinucleated giant cells, and intestinal epithelial cells. Giant cells were positive for SIV by in situ hybridization. These are the first reported cases of cryptosporidiosis with involvement of pulmonary parenchyma in SIV-infected macaques.     

Histological Characteristics of the Tracheobronchial Tree of the Least Shrew (Cryptotis Parva)

The least shrew (Cryptotis parva) is a small vomit‐competent insectivorous species which has recently been introduced as an emesis animal model in the laboratory. In this study, the respiratory system of the least shrew was examined and compared with the well‐established larger species routinely used in the laboratory. Five least shrews (4–5 g body weight, 45–60 days old) were used. Standard histological procedures were followed for light microscopic examination. The lining epithelium of the trachea was found to be pseudostratified ciliated columnar (PSCC). Three types of cells were easily identified, basal and ciliated as well as few goblet cells interspersed among the ciliated cells and they were not clearly recognizable. A few tracheal seromucous glands were located at the free end of the C‐shaped cartilaginous rings. The cartilaginous rings are replaced by smooth muscle cells before the bronchi enter into the lung. The lining epithelium of tracheobronchial tree gradually changes into simple cuboidal epithelium that lacks goblet cells. However, the division of the tracheobronchial tree is similar to other mammalian species. On the other hand, the principal bronchus lacks cartilaginous plaques as it becomes intrapulmonary bronchus. The wall of the bronchi is supported by thick layers of spirally arranged smooth muscles. Two types of cells were readily recognizable: basal and ciliated cells, with rarely observed goblet cells. In addition, the PSCC epithelium changes into simple cuboidal much earlier in the bronchial division relative to other species.     

Dose response to vaginal administration of 1,25-dihydroxyvitamin D3 to cows

It was previously reported that intravaginal (IVAG) administration of 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) might be protective against bovine hypocalcaemia. In the present study, various doses of exogenous 1,25(OH)(2)D(3) were administered IVAG to ovariectomised cows, and the subsequent changes in the biochemical parameters of the blood were measured to assess the characteristics of vaginal absorption. Five cows received 1,25(OH)(2)D(3) IVAG at a dose of 0.125, 0.25, 0.5, or 1.0microg/kg of body weight (BW) or intravenously at a dose of 1.0microg/kg BW. Dosing was at intervals of at least two weeks in a 5x5 Latin square design. Vaginally administered 1,25(OH)(2)D(3) was absorbed in a dose-dependent manner. There was no correlation between the IVAG dose of 1,25(OH)(2)D(3) and subsequent changes in plasma calcium concentrations. The bioavailability of 1,25(OH)(2)D(3) administered IVAG at 1.0microg/kg BW was approximately 93%.