On the Development of the Feline Skeleton

Light microscopic observations revealed that in camel foetuses of 25 mm crown-to-rump length (CRL) the primordial tubular system of the prospective lung was formed of several tubules lined by undifferentiated columnar epithelium. Intra-epithelial neuroendocrine cells were the first elements to be differentiated in the lining epithelium of the primordial tubular system of the prospective lung as early as 25 mm CRL. On reaching 50–67 mm CRL, the primordial tubular system started to differentiate into two systems of primordial tubules, the prospective bronchial or light tubules and the future respiratory or dark tubules. The lining epithelium of the prospective bronchial tubules revealed a clear evidence of ciliogenesis as early as 80 mm CRL. From 800 mm CRL onwards, the bronchial epithelium demonstrated ciliated and non-ciliated secretory cells. The non-ciliated secretory cells of the bronchial epithelium of fetal camel lung showed moderate reaction to AB/PAS technique, for the first time, in fetuses reaching 600 mm CRL.     

Morphological Studies on the Development of the Bronchial Epithelium of the Fetal Camel Lung

Light microscopic observations revealed that in camel foetuses of 25 mm crown‐to‐rump length (CRL) the primordial tubular system of the prospective lung was formed of several tubules lined by undifferentiated columnar epithelium. Intra‐epithelial neuroendocrine cells were the first elements to be differentiated in the lining epithelium of the primordial tubular system of the prospective lung as early as 25 mm CRL. On reaching 50–67 mm CRL, the primordial tubular system started to differentiate into two systems of primordial tubules, the prospective bronchial or light tubules and the future respiratory or dark tubules. The lining epithelium of the prospective bronchial tubules revealed a clear evidence of ciliogenesis as early as 80 mm CRL. From 800 mm CRL onwards, the bronchial epithelium demonstrated ciliated and non‐ciliated secretory cells. The non‐ciliated secretory cells of the bronchial epithelium of fetal camel lung showed moderate reaction to AB/PAS technique, for the first time, in fetuses reaching 600 mm CRL.     

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.     

Histological and histochemical study of the proventriculus (Ventriculus glandularis) of common starling (Sturnus vulgaris)

The histological and histochemical structures of the proventriculus of starling (Sturnus vulgaris) were examined using haematoxylin and eosin and special staining, that is periodic acid schiff (PAS), Masson's trichrome, Alcian blue, Orcein and Reticulin. All three cranial, middle and caudal parts of the proventriculus were also studied. The study results showed that the wall of the proventriculus consisted of mucosal, submucosal, muscular and serosal tunics. The mucosal tunic presented folds and sulci on its luminal surface. In the first third of the proventriculus, the tunica mucosa characterized by presence of folds lined by stratified squamous epithelium and presence of simple tubular glands in the lamina propria. In the middle and caudal thirds of the proventriculus, the surface was covered by a columnar epithelium, and the branched tubular glands were extended through the lamina propria. From the base of the branched tubular glands, the deep proventricular glands were observed that were compound tubuloalveolar lobules. The surface epithelium of the tunica mucosa and the cells lining the proventricular glands showed a positive reaction to PAS and Alcian blue stainings. In addition, the epithelial cells of the tubular and branched tubular glands showed Masson's trichrome-positive reaction. The submucosal tunic was thin in the proventricular wall. The tunica muscularis was formed by a thin inner layer of longitudinal smooth muscle fibres and a thick outer layer of circular fibres. The serosa consisted of loose connective tissue, rich in blood vessels and covered by mesothelium.     

Ultrastructural features of the uterus in the sexually immature ostrich (Struthio camelus) during periods of ovarian inactivity and activity

The ultrastructure of the surface epithelium and tubular glands of the uterus in the immature ostrich is described. In ostriches with inactive ovaries the uterus is lined by a non-ciliated simple columnar epithelium, with basally located heterochromatic nuclei. Scanning electron microscopy revealed that these non-ciliated cells have a dense microvillous cover. A simple columnar to pseudostratified columnar epithelium, comprised of non-ciliated and ciliated cells, lines the uterus in birds with active ovaries. The ciliated cells possess a wide luminal region, which contains a nucleus and various organelles. An accumulation of secretory granules was observed in the apical regions of the non-ciliated cells, as well as in a few ciliated cells. In addition to non-ciliated and ciliated cells, a cell type with rarefied cytoplasm was also identified. These cells appear to correspond to calcium secreting cells identified in other avian species. The results of this study indicate that, although uterine differentiation is present in immature ostriches with active ovaries, the production of secretory product appears to occur mainly in non-ciliated epithelial cells.     

Harderian Gland in Canadian Ostrich (Struthio camelus): A Morphological and Histochemical Study

Heads of ten healthy adult ostrich obtained from slaughter house were the constituted materials of the study. The Harderian gland (HG) was dissected out, and all of the gross morphometrical parameters including length, width and thickness as well as weight of left and right glands were recorded. Tissue sections were stained, using haematoxylin and eosin, Masson trichrome, periodic acid‐Schiff and Alcian blue (pH 2.5) techniques. In ostrich, HG was an orbital organ located ventromedially around the posterior part of the eyeball. It was an oval flatted shape, light pink colour with irregular outline and was pointed in the dorsal end. Its mean length was 35.30 ± 2.84 mm and 35.55 ± 3.58 mm in left and right sides, respectively, and mean width 15.30 ± 1.20 mm and 15.65 ± 1.18 mm in left and right sides, respectively. There was no significant difference between length, thickness, weight and width of left and right glands. Histological results showed that the glandular epithelium was multilobular and compound tubuloalveolar. The gland was surrounded by a connective tissue capsule, and the epithelium was lined by simple columnar epithelial cells of varying height. The secretion of HG was mucous and the secretion type was apocrine. Mucosubstance analysis revealed that secretory units contained acidic and neutral glycoproteins. The granules within the epithelial cells lining the intralobular and inter‐lobular excretory ducts of the gland were positive for periodic acid‐Schiff and Alcian blue (pH 2.5).     

THE HISTOPATHOLOGY OF INFECTIOUS BRONCHITIS IN FOWLS INFECTED WITH A NEPHROTROPIC "T" STRAIN OF VIRUS

The histopathology of the infectious bronchitis caused by the Cumming "T" strain of virus is described in fowls exposed to infection by an aerosol method. Desquamation of the ciliated and glandular epithelium throughout the trachea was seen 24 hours after exposure to virus. This was followed by rapid proliferation presumably of residual basal cells with the production of a stratified undifferentiated epithelial covering. Small areas of the tracheal submucosa showed lymphocytic infiltration by the 4th day. Cilia were first observed in the regenerating epithelium on the 7th day when mucous cells were also seen to be numerous. Alveolar mucous glands developed over the following 4 days and by the 12th day regeneration appeared complete. Pulmonary lesions were generally not severe and the air sacs were only slightly oedematous for 4 days following exposure. Necrosis of a few tubules scattered throughout the kidneys was seen on the 4th day. By the 6th day cystic tubules containing epithelial debris and polymorphonuclear leukocytes were prominent in both cortex and medulla and necrotic tubules were scattered throughout the kidneys. PAS positive granules were present in the renal tubular epithelium and were most pronounced in the distal convoluted tubules. Infiltration of the interstitium by lymphocytes and plasma cells was generally marked on the 7th day. The cytoplasm of these plasma cells was strongly PAS positive and such cells were most numerous on the 12th and 13th days after exposure and then their numbers rapidly declined. Regeneration of tubular epithelium was advanced by the 10th day and much of the cell debris had been cleared from the lumina of the tubules. What appeared to be compressed areas were seen in the cortex from the 13th day where glomeruli and tubules were numerous through considerably reduced in size. These were not seen after the 35th day, however an occasional lymph nodule persisted in the intersitium.     

The shell gland in laying and natural moulting commercial egg-type chickens: A histomorphological and ultrastructural study

The purpose of the present study was to determine the histological and ultrastructural changes in the luminal epithelium of the shell gland associated with natural moulting. Samples of the shell gland from laying (32 weeks old) and moulting (75 weeks old) hens were studied using histological, histochemical and electron microscopic techniques. In addition, TUNEL was used to demonstrate the distribution of apoptotic cells in the luminal epithelium of the shell gland. Autophagy, characterized by the presence of autophagosomes and autolysosomes, was evident in the early stages of degeneration in non-ciliated, ciliated and mitochondrial cells. The intermediate and advanced stages of regression in non-ciliated as well as mitochondrial cells occurred via apoptosis, while both apoptotic and necrotic ciliated cells were observed during the later stages of degeneration. The results of the present study suggest that a synergy of autophagy, apoptosis and necrosis is involved in the involution of the shell gland during natural moulting.     

Anatomical Features and Histology of the Nasal Cavity of One Humped Camel (Camelus dromedarius)

This study was conducted on nine heads of normal adult one-humped camels. The specimens were collected from Cairo slaughterhouse. The nasal cavity in the freshly collected samples were dissected and photographed. The specimens for microscopic studies were fixed in different fixatives and prepared to examine by light and scanning electron microscope (SEM). The nasal cavity of the camel was studied grossly and by using of light and scanning electron microscope. Specimens from different regions of this cavity were subjected to different histological stains and also demonstrated by the acid and alkaline phosphatases. Gross morphological examination of this cavity showed its three parts: rostral part (the nasal vestibule) covered with skin of usual structure then it lined with smooth mucosa. The middle part (respiratory) had dorsal, middle and ventral nasal conchae, but the caudal part (olfactory) contained the ethmoidal concha. The lining mucosa of the camel nasal cavity was similar to that of other mammals, but there were some differences: the respiratory epithelium showed a small number of goblet cells and there were a mixture of acidic and neutral mucins inside the epithelial and glandular mucous cells. Strong acid and alkaline phosphatase reaction was observed in the lining epithelium of the nasal cavity. By SEM, it showed the surface epithelial layer of the nasal cavity mucosa in three regions (vestibule, respiratory and olfactory) and resulted that it was stratified cuboidal to columnar epithelium, ciliated pseudostratified columnar epithelium with few goblet cells or olfactory mucosa containing neurosensory olfactory cells. This study aimed to investigate the anatomical features, the histological and histochemical structures of the nasal cavity in one humped camel. The findings of this study were discussed with the previous works in this field with the other domestic animals.     

A scanning electron microscopic study of the equine upper respiratory tract

The surface features of the upper respiratory tract of 20 clinically normal horses of various ages and types were studied with scanning electron microscopy. In the rostral part of the nasal cavity, there was a wide zone of non-ciliated epithelium whereas, caudally, the surface was well ciliated. This latter type of epithelium extended into the nasopharynx and guttural pouches although scattered areas of non-ciliated microvillous cells were also found.     

Isolation and monolayer culture of bovine oviduct epithelial cells

Oviduct epithelia obtained from 32 cows were cultured. The oviducts were classified into follicular and luteal phases and divided into ampulla and isthmus regions. The epithelial cells were dissociated by enzyme digestion and cultured in plastic dishes with Dulbecco's modified Eagle's medium/Ham's F12 (1:1) containing 10% calf serum. After enzyme treatment, the epithelial suspension showed free ciliated and non-ciliated cells, and cell mass. The non-ciliated cells contained secretory granules in the cytoplasm. The cell mass was composed of ciliated and secretory cells. The cell mass adhered to the dish within 12-24 hr, while the free ciliated cells attached on Day 2 of the culture. The cells grew into confluent monolayers on Day 4. The cell monolayers contained ciliated and non-ciliated cells. The monolayered non-ciliated cells showed a few secretory granules. When the cells were further cultured without subculturing, ciliary activity diminished on Day 5 and was rarely detected on Day 9. When the cells were subcultured on Day 3, ciliary movement was detected on the monolayers for only 2 days. Cell mass that did not adhere to the dish and remained floating in the medium formed ball-like structures on Day 2. Active ciliary beating was observed on the cells that were cultured in the medium supplemented with 10(-5) and 10(-9) M estradiol-17 beta, however, the ciliary activity diminished on Day 5. No difference in the cell growth was observed between the follicular and luteal phases or between the ampulla and isthmus regions.     

雏鸡哈氏腺的组织化学特点

采用常规组织化学方法对ＳＰＦ鸡的哈氏腺进行了一系列研究。结果表明，哈氏腺各级管腔上皮中有大量的杯状细胞，其分泌物主要是中性和酸性粘液物质的混合物，而腺泡上皮的分泌物主要是酸性粘液物质。改良甲苯胺蓝染色发现间质的淋巴细胞区域分布有少量的肥大细胞，管腔上皮及腺泡上皮胞浆中存在有大量异染性颗粒，预示肥大细胞在哈氏腺的局部免疫应答中起到一定的作用。苏丹黑Ｂ染色发现脂肪颗粒主要分布在上皮下。酶组织化学染色表明：各级管腔上皮及腺上皮分别为ＡＴＰ酶阳性，ＡＫＰ、ＡＣＰ及ＡＮＡＥ阴性。间质中淋巴细胞区域ＡＣＰ为强阳性，ＡＴＰ、ＡＫＰ为弱阳性，ＡＮＡＥ染色发现间质中有大量的Ｔ淋巴细胞和一定数量的巨噬细胞。此结果证明：哈氏腺是一个功能活跃的外分泌器官，Ｔ淋巴细胞和巨噬细胞在哈氏腺的局部免疫应答的调控中起到一定的作用。     

The histology and histochemical aspects of gills of the flower fish, <Emphasis Type="Italic">Pseudophoxinus antalyae</Emphasis>

Branchial epithelium of Pseudophoxinus antalyae was lined by both a thick stratified epithelium lining gill arches, gill rakers and primary filaments and a thin epithelium lining the lamellae. Mucous, chloride and rodlet cells, interspersed between pavement cells, were present in the branchial epithelium. With histochemical procedures for the characterization of glycoconjugates, mucous cells showed a strong positive reaction with Periodic acid-Shiff and Alcian Blue at pH 2.5, although with Alcian Blue at pH 0.5 and pH 1.0 the reaction was much weaker. When the combined Alcian Blue (pH 2.5) – Periodic acid-Shiff reaction was performed, most mucous cells were stained purple, whereas by the combined Aldehyde Fuchsin/Alcian Blue (pH 2.5), most cells showed a positive reaction only to Aldehyde Fuchsin. Methylation/ Alcian Blue (pH 2.5) and Methylation/ Saponification/ Alcian Blue (pH 2.5) methods showed the presence of sulphated and carboxylated glycoconjugates in mucous cells. Mucous cells were also detected to stain all metachromatically with Toluidine Blue.     

Histochemical Analysis of Glycoconjugates in the Skin of a Catfish (Arius Tenuispinis, Day)

A histochemical study using conventional carbohydrate histochemistry (periodic‐acid staining including diastase controls, alcian blue staining at pH 1 and 2.5) as well as using a battery of 14 fluorescein isothiocyanate (FITC)‐labelled lectins to identify glycoconjugates present in 10 different areas of the skin of a catfish (Arius tenuispinis) was carried out. The lectins used were: mannose‐binding lectins (Con A, LCA and PSA), galactose‐binding lectins (PNA, RCA), N‐acetylgalactosamine‐binding lectins (DBA, SBA, SJA and GSL I), N‐acetylglucosamine‐binding lectins (WGA and WGAs), fucose‐binding lectins (UEA) and lectins which bind to complex carbohydrate configurations (PHA E, PHA L). Conventional glycoconjugate staining (PAS staining, alcian blue at pH 1 and 2.5) showed that the mucous goblet cells contain a considerable amount of glycoconjugates in all locations of the skin, whereas the other unicellular gland type, the club cells, lacked these glycoconjugates. The glycoproteins found in goblet cells are neutral and therefore stain magenta when subjected to PAS staining. Alcian blue staining indicating acid glycoproteins was distinctly positive at pH 1, but gave only a comparable staining at pH 2.5. The mucus of the goblet cells therefore also contains acid glycoproteins rich in sulphate groups. Using FITC‐labelled lectins, the carbohydrate composition of the glycoproteins of goblet cells could be more fully characterized. A distinct staining of the mucus of goblet cells was found with the mannose‐binding lectins LCA and PSA; the galactosamine‐binding lectins DBA, SBA and GLS I; the glucosamine‐binding lectin WGA; and PHA E which stains glycoproteins with complex carbohydrate configurations. No reaction occurred with the fucose‐binding lectin UEA and the sialic acid‐specific lectin SNA. In addition, the galactose‐binding lectins PNA and RCA showed only a weak or completely negative staining of the mucus in the goblet cells. The specificity of the lectin staining could be proved by inhibiting binding of the lectins by competitive inhibition with the corresponding sugars. From these data, we can conclude that the mucus produced by the epidermal goblet cells of A. tenuispinis is rich in mannose, N‐acetylgalactosamine and N‐acetylglucosamine residues.     

A preliminary study on the glycosylation of the reproductive tract in the Ostrich (Struthio camelus camelus)

Glycosylation of the reproductive tract of an adult female red-necked ostrich (Struthio camelus camelus) carrying a fully formed calcified egg in her uterus when accidently killed by a blow to the head was examined using lectin histochemistry on samples from the infundibulum, magnum, uterus and vagina. Glycans in the luminal epithelium and underlying glands were described after staining with 23 lectins after neuraminidase pre-treatment in some cases. Ciliated and non-ciliated cells were evident at all levels in the luminal epithelium, the latter full of richly glycosylated secretory granules. The ciliated cells also showed glycosylation and, in the magnum, these cells often stained more intensely than the non-ciliated cells. High mannose and complex N-glycans, α1,6-linked fucosyl and sialic acid residues were present throughout the tract and there was a complete absence of GalNAcα1,3(LFucα1,2)Galß1,3/4GlcNAcß1- and rare terminal GalNAcα1- residues. Fucose in α1,2-linkage as H2 antigen and Le^y was also rare in the luminal epithelium and completely absent in glands. Terminal galactose was present in the luminal epithelium apart from in the infundibulum. Gland epithelium showed similar glycosylation to the luminal epithelium except in the magnum where there were significant differences and here the glands were packed full of large secretory granules, unlike the glands in the rest of the tract. Each section of the tract had its own specific pattern of glycosylation which could be related to the stage of egg formation.     

Staining pattern of the brush border and detection of cytoplasmic granules in the uriniferous tubules of female DBA/2Cr mouse kidney: comparison among various fixations and stains.

The proximal straight tubules of the female mouse kidney exhibit heavy periodic acid Schiff (PAS) staining in their brush borders and numerous cytoplasmic granules. In the present study, the female DBA/2Cr mouse kidney was examined, using various fixatives (formalin, PFA, PLP, Zamboni's, Bouin, or Carnoy solution) and various staining methods (HE, PAS, alcian blue, periodic acid methenamine-silver (PAM), toluidine blue, azan, or Congo red). Under azan and PAM, the staining pattern of the brush border was similar to that of PAS, and few effects of the fixative were observed. Cytoplasmic granules were clearly detected with PAM as well as PAS. However, these granules were not detected with Carnoy solution. Furthermore, distribution of granules differed between PAS and PAM.     

Uptake of ferritin and Bordetella avium in bronchus-associated lymphoid tissue of turkeys

The uptake of macromolecular and particulate materials in bronchus-associated lymphoid tissue (BALT) in turkeys was examined using transmission electron microscopy. Tracer materials used were live and ultraviolet-killed (UV-killed) Bordetella avium and ferritin. Suspensions of bacteria and ferritin were instilled via intratracheal catheterization and allowed to remain in contact with the respiratory surfaces for 0, 10, 30, 60, 90, and 120 min. Ferritin and B. avium were taken up by both ciliated and non-ciliated cells of the epithelium overlying BALT (BALT epithelium). Ferritin was found in organelles associated with endocytosis (i.e. apical vesicles, endosomes, cytoplasmic vacuoles) and was apparently transported across epithelial cells, since it was also found in intercellular spaces. Bacteria were found in vacuoles within BALT epithelial cells, but not free in intercellular spaces. Some macrophages in BALT epithelium also contained bacteria. No differences were observed between uptake of live and UV-killed bacteria. We conclude that both ciliated and non-ciliated cells of BALT epithelium in turkeys are able to take up macromolecular and particulate materials. Bacteria are also accessible to intraepithelial macrophages, although whether they are taken up directly from the bronchial surface or whether they pass through epithelial cells first could not be determined. This evidence suggests that antigens, including respiratory pathogens, could gain access to cells of the avian immune system by transepithelial passage in BALT.     

Epithelial cell kinetics in the inflammatory process of chicken trachea infected with infectious bronchitis virus

All stages of degeneration and regeneration in chicken tracheal epithelium were studied morphologically following an intratracheal inoculation of infectious bronchitis virus (IBV). Viral antigen was detected in the cytoplasm of tracheal epithelium from 1 to 7 days post-inoculation (d.p.i.) with a peak on 3 d.p.i. At 1 d.p.i., almost all epithelial cells were involved in the degeneration. At this time, labelling index of bromodeoxyuridine (BrdU) in the basal cells showed significantly high value compared with control. At 2 and 3 d.p.i., a great number of basal cells were recognized, but the BrdU labelling index tended to decrease. At 4 and 5 d.p.i., the BrdU labelling index of basal cells significantly decreased than 1 d.p.i., and a few number of regenerated immature ciliated epithelia appeared. At 6 to 11 d.p.i., the ciliated columnar epithelia increased rapidly in number, and returned to the normal appearance except for non-ciliated patch by 13 d.p.i. These results suggested that the tracheal epithelial cells infected with IBV degenerated within 24 hours and proliferating activity of basal cells functioned immediately, and 3 to 4 days later, these basal cells were differentiated to the ciliated epithelia.     

Normal ultrastructure and histochemical characteristics of canine lacrimal glands

Lacrimal glands of 12 dogs free of ocular disease were examined to determine the normal structure of these glands. The glands consisted of tubuloacinar cells that ultrastructurally and histochemically were of a single type of secretory cell in the tubules and possibly 3 types of secretory cells in the acini. The tubular epithelium contained homogenous electron-dense granules that stained as neutral glycoconjugates (periodic acid-Schiff positive and Alcian blue and high iron diamine negative). The predominant acinar cells contained granules of lesser electron density than those of the tubules, and stained as sialomucin (Alcian blue [pH 2.5] and periodic acid-Schiff-positive, and high iron diamine-negative). A second type of acinar cell was in peripheral lobules that ultrastructurally and histochemically appeared like lipid granules (positive with oil red O and osmium tetroxide). Ultrastructurally, a third type of acinar granule was finely granular, electron-lucent, and frequently coalesced. It was not readily apparent whether the latter was an artifact, a stage in the maturation of the sialomucin granules, or a third type of acinar granule. Individual acinar cells usually had a predominance of 1 granule type, but greater than 1 granule type could be found in some cells. The basal surfaces of the acinar, tubular, and ductal cells were incompletely ensheathed by myoepithelial cells. Plasma cells, lymphocytes, mast cells, endothelial cells, fat cells, and Schwann cells composed the cellular elements of the interstitium. Lymphocytes, mast cells, and nerve endings also were found in the parenchyma.