Morphological Studies on the Harderian Gland in the Ostrich (Struthio camelus domesticus) on the Embryonic and Post‐natal Period

The present investigation was performed on 50 ostriches from 28th day of incubation until the 7th month of life. The morphological (morphometric, histological, histometric and histochemical) studies were conducted. Tissue sections were stained with haematoxylin–eosin, methyl green‐pyronin Y, periodic acid–Schiff, alcian blue pH 2.5, aldehyde fuchsin and Hale's dialyzed iron studies. The Harderian gland becomes macroscopically visible on the 28th day of incubation. It is situated in the ventronasal angle of the orbit near inter‐orbital septum, between medial rectus muscle, pyramidal and ventral oblique muscles. The Harderian gland of ostrich is a tubulo‐acinar gland. The acini were composed of tall conical cells which formed a small lumen and were surrounded by myoepithelial cells. These cells had a granular basophilic, vacuolated cytoplasm. Each of the lobes has a system of complex branching ducts – tertiary, secondary and primary. In the III of research group (3rd week of life), the presence of few plasma cells was demonstrated, which were located within acini and tertiary and secondary ducts, whereas the biggest concentration of plasma cells was observed in group IV of research tissue (4th month of life). The dark cells were observed first time in main ducts 72 h after hatching of nestlings (group II). The morphometric and histometric studies showed that the most intensive growth of Harderian gland occurred between the third week and the seventh month of birds' life. The histochemical study indicated the presence of neutral and acidic mucins, glycoproteins and carboxylated acid mucopolysaccharides.     

Expression of different classes of immunoglobulin in intraepithelial plasma cells of the Harderian gland of domestic ducks Anas platyrhynchos

The Harderian gland of chickens contains numerous plasma cells and is considered as a peripheral lymphoid organ. Data about this gland in other avian species are scarce or inexistent. Considering that ducks show some unique characteristics regarding the immune system, which are important in evolutionary context, and that unusual location of plasma cells into the epithelium was recently described in primitive avian species, here we investigated the occurrence and characterized intraepithelial plasma cells in the Harderian gland of ducks, according to the immunoglobulin produced. Numerous intraepithelial plasma cells were found confined to the Harderian gland ducts. Plasma cells were also found in the ducts lamina propria. IgM-positive cells were the most abundant into the epithelium. In contrast, IgY- or IgA-positive cells were predominant in the lamina propria. The constancy of intraepithelial plasma cells in all specimens examined indicates that they may be essential mediator for an effective immunesurvaillance of the ocular mucosa.     

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).     

Normal anatomical and histochemical characteristics of the lacrimal glands in the American bison and cattle

Dorsal lacrimal glands, superior glands of the third eyelid and Harderian glands (deep gland of the third eyelid) from 19 bison and 18 cattle free of apparent ocular disease were examined to compare the normal anatomical properties of these glands. All glands were characterized and measured (length and width). The gross anatomy of the dorsal lacrimal glands was similar, with the exception of a bipartite gland in cattle. The bison's superior gland of the third eyelid and Harderian gland was longer as compared with cattle. A subset of the bison and cattle samples (five bison and five cattle) was sectioned for histological and histochemical analysis. The histology of the dorsal lacrimal and superior gland of the third eyelid revealed tubuloalveolar cells with basophilic vacuolated cytoplasm in bison and eosinophilic granular cytoplasm in cattle. The Harderian glands consisted of a tubuloalveolar anterior part combined with large lumens acini lined with cuboidal epithelium in the posterior part; the posterior part of the bison Harderian gland was more predominant than in cattle samples. Mucosubstance histochemistry revealed acidic and neutral glycoproteins with similar staining patterns in all glands of both species.     

Histological and immunological studies on the fowl lacrimal gland following surgical excision of Harder's gland.

Surgical removal of the Harderian gland of the domestic fowl resulted in increased secretory activity in the lacrimal gland and also in an increase in goblet cell numbers along the length of the lacrimal gland duct. Plasma cells were more numerous in the lacrimal glands of operated birds and they were capable of antibody responses to both systemically and topically applied bovine serum albumin (BSA). Gel diffusion studies showed the presence of anti-BSA activity in the tears and serum of fowls stimulated after surgical removal of the Harderian gland.     

Spontaneous Harderian Gland Adenocarcinoma in a Female F344 Rat: A Case Report

Harderian gland tumors are extremely rare in female F344 rats. An expansive enlarging lesion of the Harderian gland with compression, distortion and invasion of the surrounding muscle was found in a 110-week-old female F344/DuCrj rat, which was diagnosed as a Harderian gland adenocarcinoma. Epithelial growth patterns such as glandular, lobular, papillary and duct forming patterns were exhibited in most areas of the tumor. The tumor cells were pleomorphic and atypical. In one part of the tumor, poorly differentiated areas were found. This case was observed in the middle dose group of a carcinogenicity study of diphenylamine, which was not carcinogenic, we determine to be this case was a spontaneous tumor.     

Effect of Harderian adenectomy on the statistical analyses of mouse brain imaging using positron emission tomography

Positron emission tomography (PET) using 2-deoxy-2-[¹⁸F] fluoro-D-glucose (FDG) as a radioactive tracer is a useful technique for in vivo brain imaging. However, the anatomical and physiological features of the Harderian gland limit the use of FDG-PET imaging in the mouse brain. The gland shows strong FDG uptake, which in turn results in distorted PET images of the frontal brain region. The purpose of this study was to determine if a simple surgical procedure to remove the Harderian gland prior to PET imaging of mouse brains could reduce or eliminate FDG uptake. Measurement of FDG uptake in unilaterally adenectomized mice showed that the radioactive signal emitted from the intact Harderian gland distorts frontal brain region images. Spatial parametric measurement analysis demonstrated that the presence of the Harderian gland could prevent accurate assessment of brain PET imaging. Bilateral Harderian adenectomy efficiently eliminated unwanted radioactive signal spillover into the frontal brain region beginning on postoperative Day 10. Harderian adenectomy did not cause any post-operative complications during the experimental period. These findings demonstrate the benefits of performing a Harderian adenectomy prior to PET imaging of mouse brains.     

网状内皮增殖病病毒(REV)感染SPF雏鸡后局部免疫组织抗体生成细胞的变化

对 SPF雏鸡人工感染 REV后局部免疫组织盲肠扁桃体和哈德氏腺的 Ig A、Ig G和 Ig M抗体生成细胞数量动态变化进行了检测。结果发现 ,REV感染 SPF雏鸡后 7～ 4 9天消化道局部免疫组织盲肠扁桃体和呼吸道相关局部免疫组织哈德氏腺的 Ig A、Ig G和 Ig M抗体生成细胞数量明显低于未感染对照雏鸡。表明雏鸡感染 REV后消化道局部组织盲肠扁桃体和呼吸道相关免疫组织哈德氏腺体液免疫机能均显著下降。     

Local immune response in the chicken Harderian gland to antigen given by different ocular routes.

The effectiveness of three ocular routes of antigen administration to produce a local immune response in the Harderian gland was studied. The routes were by eyedrop, injection into the ocular conjunctiva and injection into the nictitating membrane. The antigen was observed in the cytoplasm of macrophages located within the lymphoid tissue only after the injection into the nictitating membrane. The numbers of germinal centres and plaque forming cells found in the gland after injection into the nictitating membrane was higher than the numbers observed following the other two ocular applications. These findings indicate that the injection of the antigen into the nictitating membrane is the most effective ocular route for producing a local immune response in the Harderian gland.     

Scanning Electron Microscopy of the Orbital Harderian Gland in the Male Atlantic Bottlenose Dolphin (Tursiops truncatus)

The ultrastructure of the Harderian gland of Atlantic bottlenose dolphin ( Tursiops truncatus ) was examined by scanning electron microscopy (SEM). We found the following surface features: the typical round appearance of the ascinar glandular unit with a finely granular surface, a thin cortex and immediately below two types of cells: type I cells (characterized by small lipid vacuoles) and type II cells (characterized by large lipid vacuoles). It has been suggested that different cells forms represent a single cell type in varying activity states. Additionally, a coalescent tubular complex, a small balloon-like structures and large globular structures were observed. These structures may be reservoirs of secretion products.     

鸡贫血因子病局部粘膜免疫功能变化的研究

用鸡贫血病毒感染１日龄ＡＡ雏鸡,以未感染同龄ＡＡ雏鸡的对照,在感染后７,１４,２１,２８,３５,４２,４９日龄检测其哈德尔腺及盲肠扁桃体Ｔ细胞和ＩｇＧ＾＋、ＩｇＭ＾＋、ＩｇＡ＾＋抗体生成细胞数量,泪液,气管液、肠液中ＩｇＭ、ＩｇＭ、ＩｇＡ含量的动态变化。     

A morphological and immunohistochemical study of healthy and atretic follicles in the ovary of the sexually immature ostrich (Struthio camelus)

The morphology of healthy and atretic follicles in the ovary of the sexually immature ostrich was described in the present study. In addition, the distribution of the intermediate filaments desmin, vimentin and smooth muscle actin, in these ovarian follicles, was demonstrated. Healthy and atretic primordial, pre-vitellogenic and vitellogenic follicles were present in the ovaries of the sexually immature ostrich. Atresia occurred during all stages of follicular development. Atretic primordial and pre-vitellogenic follicles were characterized by the presence of a shrunken oocyte surrounded by a multilayered granulosa cell layer. Two forms of atresia (types 1 and 2) were identified in vitellogenic follicles. In the advanced stages of type 1 atresia the follicle was dominated by a hyalinized mass. In contrast, in type 2 atresia the granulosa and theca interna cells differentiated into interstitial gland cells. Positive immunostaining for desmin was observed in the granulosa cells of only healthy primordial and pre-vitellogenic follicles. Atretic primordial and pre-vitellogenic follicles were immunonegative for desmin. Vimentin immunoreactivity was demonstrated in the granulosa cells of all follicles except the vitellogenic atretic follicles. The results of the present study indicate that ovarian follicles in the sexually immature ostrich undergo a cycle of growth and regression, which is similar to that reported in other avian species. Furthermore, based on the results of the immunohistochemical study, it would appear that the distribution and immunostaining of intermediate filaments changes during follicular development and atresia.     

切除结膜结合淋巴组织鸡对新城疫疫苗点眼免疫反应 Ⅱ.组织学与免疫组织化学研究

60只 1日龄健康雏鸡随机分为 A、B、C三组。A组于 1日龄手术切除下眼睑结膜。A、B两组于 12日龄时结扎鼻泪管 ,用新城疫克隆 30苗点眼。C组为对照组。 2 3日龄分别采取其 CAL T和哈德氏腺 ,比较观察其组织细胞。结果 ,1日龄手术切除下眼睑结膜后造成鸡 CAL T缺失 ,点眼免疫后 ,哈氏腺的免疫细胞也较正常免疫鸡少 ,而正常接种 ND克隆 30 (B组 )后 ,CAL T发生早 ,生长快 ,淋巴细胞数量多 ,故本实验证实 CAL T对哈氏腺免疫细胞的数量与成分有一定的调控作用。 CAL T缺失鸡是研究黏膜免疫系统比较理想的模型     

切除结膜结合淋巴组织鸡对新城疫疫苗的免疫反应Ⅰ.局部与循环抗体检测

实验将 60只 1日龄健康雏鸡随机分为A、B、C 3组。A组为CALT缺失组 ,B组为免疫组 ,C组为非免疫对照组。A组于 1日龄手术切除下眼睑结膜 ,A、B两组于 1 2日龄结扎鼻泪管后进行点眼免疫接种新城疫克隆 30疫苗。2 3日龄时 ,收集A、B、C各实验组鸡的血清和泪液 ,做红细胞凝集抑制试验 (HI试验 )、酶联免疫吸附试验(ELISA)。比较观察各实验组雏鸡血清和泪液抗体HI效价与IgA、IgG、IgM含量的变化特点。结果 ,1日龄手术切除下眼睑结膜后造成鸡CALT缺失 ,CALT缺失鸡点眼免疫后 ,泪液抗体HI效价与IgA、IgG、IgM含量比免疫组雏鸡低。但对雏鸡的抗体生成的规律并不产影响。故实验证实了CALT对眼区局部抗体的产生及其种类有重要的调控作用 ,协同哈氏腺完成局部免疫的功能     

Investigations on the conjunctival goblet cells and on the characteristics of glands associated with the eye in the guinea pig

Objective To investigate the distribution and density of conjunctival goblet cells (GC) and to study the anatomy and microscopic characteristics of glands associated with the eye in the guinea pig. Procedures Twenty‐five guinea pigs were used. Meibomian gland openings were counted using biomicroscopy. Conjunctiva, eyelids and glands were embedded in glycol methacrylate and paraffin. Sections were stained with hematoxylin and eosin (H&E), periodic acid Schiff’s reaction (PAS) and Alcian blue (AB). Results Highest GC densities were found in the bulbar and palpebral region of the nasal conjunctiva (GC index: 13.7–16.4%). Lowest GC densities (GC index: 0.0–1.0%) were found in 3/4 limbal regions (nasal and temporal upper eyelid, temporal lower eyelid). Guinea pigs have 27.1 ± 3.0 (mean ± SD) meibomian gland openings in the upper lid and 25.7 ± 2.3 in the lower lid. Difference between upper and lower lid was significant (P = 0.037). Two subconjunctival sebaceous glands occur temporal to each eye. The Harderian gland is very large. In the lacrimal gland three different cell types were distinguished both according to the cell structure and histochemical staining. Conclusions Goblet cell densities are lower in guinea pigs than in dogs and horses. Positive staining with PAS and AB could be an indication that mucins are produced in the lacrimal gland. If so, they may contribute to the mucin layer of the tear film. Both the extraordinarily large Harderian gland and the subconjunctival sebaceous glands produce lipids and may contribute to the lipid layer of the tear film.     

Further observations on functional deletion of paraocular glands in the fowl (Gallus domesticus)

Methods are described for inoculation of the fowl Harderian gland with sclerosing reagent and for its surgical excision. Though very destructive the former procedure proved the less reliable in achieving the gland's complete functional deletion. Three weeks after surgical removal of the Harderian glands the lachrymal glands of 10-week-old fowls were heavier and contained more immunocompetent cells than the glands of intact and sham operated birds. When adult birds deprived of both paraocular glands were given sheep erythrocytes or Newcastle disease virus by eye drop they developed slightly higher than normal titres of serum antibody but failed to produce lachrymal antibody.     

The orbital Harderian gland of the male atlantic bottlenose dolphin (Tursiops truncatus): a morphological study

The ultrastructure of the Atlantic Bottlenose dolphin Harderian gland (HG) has been described but some questions remain unanswered. The purpose of this work was to define the gland's structure, ultrastructure and the differences between cells (types I and II) of the male dolphin using optic, fluorescence and electron transmission microscopy. Three different cells were observed under optic and fluorescence microscopic examination, while only two cell types (types I and II) were distinguished by electron transmission microscopy. Type I (oval nuclear envelope) exhibited three different cell populations and type II (indented nuclear envelope) exhibited two different cell populations. Although, we observed both types of vesicles in both types of cells they differed, principally, in quantity. The glands also possessed prominent duct systems, with three orders of complexity. The dolphin orbital HG appears to function as a mixed heterologous gland with two types of cells that exhibit both types of vesicles and other distinguishable differences.     

The Excurrent Ducts of the Testis of the Emu (Dromaius novaehollandiae) and Ostrich (Struthio camelus): Microstereology of the Epididymis and Immunohistochemistry of its Cytoskeletal Systems

The volumetric proportion of the various ducts of the epididymis of the emu and ostrich and the immunohistochemistry of actin microfilaments, as well as cytokeratin, desmin and vimentin intermediate filaments, were studied in the various ducts of the epididymis of the emu and ostrich. The volumetric proportions of various ducts, which are remarkably different from those of members of the Galloanserae monophyly, are as follows: the rete testis, 5.2 ± 1.4% for the emu and 2.4 ± 1.8% for the ostrich; efferent ducts, 14.2 ± 2.3% (emu) and 11.8 ± 1.8% (ostrich); epididymal duct unit, 25.8 ± 5.8% (emu) and 26.1 ± 4.1% (ostrich) and connective tissue and its content, 54.7 ± 5.8% (emu) and 60.0 ± 4.9% (ostrich). Unlike in mammals and members of the Galloanserae monophyly, only vimentin was immunohistochemically demonstrated in the rete testis epithelium of the emu, and none of the cytoskeletal protein elements in the ostrich rete testis. The epithelium of the efferent ducts of the emu co-expressed actin, cytokeratin and desmin in the non-ciliated type I cells, and vimentin in the ciliated cell component. The ostrich demonstrated only cytokeratin in this epithelium. The ratite epididymal duct unit is different from that of mammals in lacking actin (only weaky expression in the ostrich), desmin and cytokeratin, and a moderate/strong immunoexpression of vimentin in the basal cells and basal parts of the NC type III cell in the epididymal duct unit. Immunoexpression of the microfilaments and intermediate filaments varied between the two ratite birds, as has been demonstrated previously in birds of the Galloanserae monophyly, and in mammals.     

Different patterns in the histology and autofluorescence of the Harderian glands of the Syrian Hamster, rat, mouse, Mongolian gerbil and guinea pig

The purpose of this study was to compare the natural fluorescence in the Harderian glands of the Syrian hamster, rat, mouse, Mongolian gerbil and guinea pig (both sexes). For each species, 10 animals (five males and five females) were used. Histological autofluorescence studies were performed using a fluorescence microscope (450-490 nm filter). Two different types of fluorescent cells were observed in both hamster (type AFI high intensity and type AFII, low fluorescence) and rat (type AFI, low fluorescence and type AFII, high fluorescence) Harderian glands. The fluorescence was basally located in all mice cells, whereas it was observed near the epithelial cell nuclei in the Mongolian gerbil (occupying two-thirds and one-third of the cells in males and females, respectively). A high intensity of fluorescence was present throughout the acinar cells in the guinea pig. The patterns of fluorescence identified exhibited a sexual dimorphism in all species studied. These results demonstrate that the Harderian glands of the animal species examined exhibit a variety of histological autofluorescence patterns.     

Investigations on the conjunctival goblet cells and the characteristics of the glands associated with the eye in chinchillas (Chinchilla Laniger)

Objective To investigate the density and distribution of conjunctival goblet cells (GC) and study the anatomy and microscopic characteristics of glands associated with the eye in chinchillas (Chinchilla Laniger). Procedure 12 chinchillas were included in the study. Conjunctiva (divided into four regions), eyelids, and glands were embedded in paraffin wax, sectioned, stained, and analyzed. Results Highest GC densities were found in the palpebral region of the nasal and temporal conjunctiva of both eyelids (GC index: 25.1-18.2%), and lowest densities, in the bulbar and marginal region of the nasal and temporal conjunctiva of both eyelids (GC index: 1.5-0.0%). Meibomian glands extend along the entire length of both eyelids, and the whole glandular complex broadens toward the temporal canthus. This is macroscopically visible through the conjunctiva. The openings of the Meibomian glands are macroscopically not discernible. The light pink, smooth, and crescent-shaped lacrimal gland lies next to the aforementioned broadened part of the Meibomian glands in the temporal canthus. The whitish, 0.9-cm-long, smooth Harderian gland is firmly attached to the posterior part of the globe and extends nasally from the optic nerve to the equator. Furthermore, chinchillas possess two lacrimal puncta, situated on the inner conjunctival surface of both eyelids near the medial canthus. A pigmented lacrimal canaliculus originates from each punctum. The vestigial nictitating membrane is supported by a hyaline cartilage and is pigmented at its free margin. Conclusions Chinchillas possess a Harderian gland, a lacrimal gland, and Meibomian glands. The GC density in the nasal and temporal palpebral conjunctiva is higher than in guinea pigs.