Macroscopic and microscopic analysis of the tongue of the common opossum (Didelphis marsupialis)

We performed a macroscopic and microscopic study of the tongues of common opossums, Didelphis marsupialis, from South America. We studied two males and two females. We collected morphometric data on the tongue with precision calipers. For the light microscopy and scanning electron microscopy analyses, we fixed tissue fragments in 10% formaldehyde and 2.5% glutaraldehyde, respectively. The opossum tongues averaged 5.87 ± 0.20 cm in length, 3.27 ± 0.15 cm in width at the lingual body, and 3.82 ± 0.15 cm in width at the root. The mean thickness of the lingual body was 1.8 ± 0.1 cm, and the thickness of the root was 3.82 ± 0.15 cm. Sharp filiform papillae were scattered across the entire tongue; conical filiform papillae occurred on the lingual body and tongue tip; fungiform papillae were scattered among the filiform papillae on the lingual body and tongue tip; and there were three vallate papillae at the root of the tongue. We found two strands of papillary projections in the tongue root. Despite the low variability observed in the lingual papillae, the morphological data obtained in this study may be related to the opossum's diverse food habits and the extensive geographic distribution of the species throughout America. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

Morphofunctional structure of the lingual papillae in three species of South American Camelids: Alpaca,guanaco, and llama

The aim of this study was to compare the anatomical and functional characteristics of the lingual papilla among the Camelidae. For this purpose, tongues of alpaca, guanaco, and llama were used. Numerous long and thin filiform papillae were located in the median groove and none were detected on the rest of the dorsal surface of the lingual apex in alpaca. Secondary papillae originated from the base of some filiform papillae on the ventral surface of alpaca tongue. The bases of some filiform papillae of the lateral surface of the lingual apex were inserted into conspicuous grooves in guanaco and tips of filiform papillae on the dorsal surface of the lingual body were ended by bifurcated apex. On the dorsal surface of the lingual apex of llama, there were no filiform papillae but there were numerous filiform papillae on both the lateral margins of the ventral surface of the lingual apex. Fungiform papillae were distributed randomly on dorsal lingual surface and ventral margins of the tongues of all camelid species. Lenticular papillae were located on the lingual torus and varied in size and topographical distribution for each species. Circumvallate papillae had irregular surfaces in llama and alpaca, and smooth surface in guanaco. In conclusion, llama and alpaca tongues were more similar to each other, and tongues of all camelid species displayed more similarities to those of Bactrian and dromedary camels in comparison with other herbivores and ruminants. Microsc. Res. Tech. 79:61–71, 2016. © 2015 Wiley Periodicals, Inc.     

Macro‐ and microscopic study on the tongue and lingual papillae of Bison bonasus hybrid as an interspecific species (Bos taurus × Bison bonasus)

Nowadays the processes of crossing of different mammalian species are well characterized in light of genetics, and possibility of reproduction. Generally, lack detailed studies on changes in anatomy and histology of internal systems of hybrids. Bison bonasus hybrid is an interspecific species between the bull of European bison (Bison bonasus), and female of domesticated cattle (Bos taurus). To study the anatomy of tongue and distribution of lingual papillae on Bison bonasus hybrid tongue, a stereomicroscopic and scanning electron microscopic observations was conducted. We aimed to study the appearance of macro‐ and microscopic traits in relation to parental species and other bovids. Results indicate that Bison bonasus hybrid tongue possesses characteristic traits for parental species and ruminants from Bovidae family and also species‐specific features differ from mentioned animals. Specific new traits in hybrid are: V‐shaped arrangement of lingual papillae on ventral surface of the tongue, rosette arrangement of conical papillae on the top of lingual prominence, bigger number of vallate papillae and closer arrangement of vallate papillae. In hybrid observed also the appearance of combination of parental traits. Similar to European bison are rectangular shape of lingual prominence and absence of filiform papillae on the root of tongue. Presence of lateral processes of filiform papillae on the apex of tongue, and arrangement of vallate papillae into two rows corresponded to cattle. Development of new morphological features of tongue and its mucosal papillae in Bison bonasus hybrid opens new issue for future studies on organs of digestive system in animals after hybridization.     

Histatin-induced alterations in Candida albicans: a microscopic and submicroscopic comparison

Despite the numerous studies performed in an attempt to clarify the issue, the mechanism of action of salivary histatins remains unclear. The aim of the present study was to correlate histatin-induced morphological changes in Candida albicans by fluorescence microscopy (FM), transmission electron microscopy (TEM), and high resolution scanning electron microscopy (HRSEM). Each of the fluorescent dyes used by FM (i.e., tetramethylrhodamine methyl ester perchlorate for mitochondrial potential, Lysotracker for lysosome acidic compartment, and 4',6-diamino-2-phenylindole dihydrochloride for DNA) exhibited a specific staining in control cells. Following histatin treatment, we observed a recurring staining pattern, corresponding to fluorescence concentration along the cell periphery, suggesting a loss of dye specificity. To assess histatin-induced cytoplasmic modifications, ultrastructural analysis was then carried out. After treatments with histatins, TEM revealed characteristic intracellular modifications including: vacuole overgrowth, nuclear disappearance, loss of organelle identity, as well as the appearance of electron-dense membranes, likely of mitochondrial origin. Additionally, structures resembling autophagosomes were occasionally observed. By HRSEM, mitochondrial swelling was invariably the first sign of a histatin-induced effect. Other modifications included intracellular membrane disarrangement, organelles in disarray, and a large central cavity with deformed bodies displaced to the cell periphery, similar to what was detected by TEM. In summary, our study illustrates the occurrence of ultrastructural modifications following administration of histatins. Observations made with FM, TEM, and HRSEM provided different views of the same signs, demonstrating a definite action of histatins on C. albicans morphology. The possible functional meanings of these morphological results is discussed in light of the most recent biochemical data on histatin fungicidal activity.     

Morphology of the lingual surface of South American fur seal (Arctocephalus australis) and sea lion (Otaria flavescens)

In this study, we aimed to describe the morphological characteristics of the lingual papillae in two species of Otariidae family by stereomicroscopy and scanning electron microscopy. We used tongues of two South American Otariidae species. The tongues were elongated and terminated in bifid apex and there was no median sulcus on the dorsal lingual surface. The most numerous type of lingual papilla was filiform in the South American fur seal (SASL) and entire dorsal lingual surface was covered by these filiform papillae but the dorsal surface of the tongue of the South American sea lion was covered by numerous polygonal projections, which were different in size. Fungiform papillae were detected in only SASL and they randomly distributed on the lingual apex and body, and some fungiform papillae were collected into twosome or threesome groups on the posterior part of the lingual body. Circumvallate papilla was found in the center of the lingual radix of South American sea lion. Thread‐like conical papillae were common for both species and they located on the lingual radix. We determined that lingual surface morphology was completely different in each species, although they were members of the same family, Otariidae. Microsc. Res. Tech. 78:140–147, 2015. © 2014 Wiley Periodicals, Inc.     

Ultrastructure of submandibular salivary glands of mouse: TEM and HRSEM observations

The fine structure of submandibular glands of mouse were analyzed using light microscopy (LM), high resolution scanning electron microscopy (HRSEM), and transmission electron microscopy (TEM) methods. For LM, the specimens were embedded in Spurr resin, stained by toluidin blue solutions. For TEM, the tissues of submandibular salivary glands were fixed with modified Karnovsky solution and postfixed with osmium tetroxide. For HRSEM, the tissues were fixed with 2% osmium tetroxide solution in 1/15M sodium phosphate buffer (pH 7.4). The samples were immersed successively in dymethylsulphoxide and freeze cracked. The maceration was made in diluted osmium tetroxide for 24-48 h. The samples were examined by high resolution scanning electron microscopy. The intracellular components of acinar and ductal cells revealed clearly the Golgi apparatus, rough endoplasmic reticulum, secretory granules, and mitochondria. The end bulbs of Golgi lamellae and flattened cisterns of rough endoplasmic reticulum showed the luminal surface. A few mitochondria were identified intermingling between the rough endoplasmic reticulum and the mitochondriales cristae in three-dimensional HRSEM images. Secretory granules were numerous and presented different sizes. Small granules of ribosomes were attached on cistern surface, measuring 20-25 nm in diameter. Numerous arranged microvilli were found on the luminal surface of secretory canaliculus. The contact surfaces of acinar cells revealed complicated interdigitations by cytoplasmic processes. The mitochondria of duct cells were disposed vertically and surrounded by basal infoldings of plasma membranes. Basement membrane showed a spongy-like structure having an irregular surface with various strands and meshes of fine collagen fibrils.     

The fine structure of fenestrated adrenocortical capillaries revealed by in-lens field-emission scanning electron microscopy and scanning transmission electron microscopy

Cell biologists probing the physiologic movement of macromolecules and solutes across the fenestrated microvascular endothelial cell have used electron microscopy to locate the postulated pore within the fenestrae. Prior to the advent of in-lens field-emission high-resolution scanning electron microscopy (HRSEM) and ultrathin m et al coating technology, quick-freeze, platinum-carbon replica and grazing thin-section transmission electron microscopy (TEM) methods provided two-dimensional or indirect imaging methods. Wedge-shaped octagonal channels composed of fibrils interwoven in a central mesh were depicted as the filtering structures of fenestral diaphragms in images of platinum replicas enhanced by photographic augmentation. However, image accuracy was limited to replication of the cell surface. Subsequent to this, HRSEM technology was developed and provided a high-fidelity, three-dimensional topographic image of the fenestral surface directly from a fixed and dried bulk adrenal specimen coated with a 1 nm chromium film. First described from TEM replicas, the “flower-like” structure comprising the fenestral pores was readily visualized by HRSEM. High-resolution images contained particulate ectodomains on the lumenal surface of the endothelial cell membrane. Particles arranged in a rough octagonal shape formed the fenestral rim. Digital acquisition of analog photographic recordings revealed a filamentous meshwork in the diaphragm, thus confirming and extending observations from replica and grazing section TEM preparations. Endothelial cell pockets, first described in murine renal peritubular capillaries, were observed in rhesus and rabbit adrenocortical capillaries. This report features recent observations of fenestral diaphragms and endothelial pockets fitted with multiple diaphragms utilizing a Schottky field-emission electron microscope. In-lens staging of bulk and thin section specimens allowed tandem imaging in HRSEM and scanning TEM modes at 25 kV.     

The structure of the developing chick retinal pigment epithelium revealed by high resolution scanning electron microscopy

The retinal pigment epithelium (RPE) in the developing eye of chick embryos has been studied during the early stages of development by high resolution scanning electron microscopy (HRSEM). Specimen preparation techniques which involve removal of the cytoplasmic matrix permitted visualization of organelles and other subcellular structures within RPE cells in detail and in three dimensional (3-D) stereo HRSEM. Using this technique, we were able to examine changes in melanosome structures during development and demonstrate that pigmentation in the RPE was present by day 4 of development. RPE plasma cell membranes showed extensive folding of the apical portion of the membrane closest to the developing neural retina by day 9. Examination of RPE photoreceptor junction revealed photoreceptor inner segments by day 6 and an outer segment by day 9. Mitochondria in the RPE were found to contain tubular cristae only. The ultrastructure in 3-D of the Golgi apparatus, smooth and rough endoplasmic reticulum, lysosomes and nuclear chromatin of the RPE, and Bruch's layer was revealed by the HRSEM method.     

Electron microscopic observations of stomata,epicuticular waxes,and papillae in Chamaecyparis obtusa: Reconsidering the traditional concept of Y‐shaped white stomatal bands

The foliar morphological characters of hinoki (Chamaecyparis obtusa) were revisited using optical and scanning electron microscopy. In C. obtusa, typical Y‐shaped white stomatal bands were evident on the abaxial leaf surfaces. Two facial leaves and two lateral leaves were observed at the same node. Waxy papillae and oval stomata were arranged in two or three rows with protuberant rims on the abaxial leaf surfaces. Higher magnifications revealed the deposition of epicuticular waxes (tubules) on the Y‐shaped white stomatal bands. Given the absence of stomatal bands after dewaxing with organic solvents, the white stomatal bands in C. obtusa were related to the epicuticular waxes rather than the presence of aggregated stomata alone. In contrast to C. obtusa, a single median leaf and two lateral leaves were observed at the same node of oriental arborvitae (Platycladus koraiensis). Neither stomatal bands nor papillae were observed on P. koraiensis leaves. The stomatal density and epicuticular waxes in the stomatal regions of C. obtusa were higher than those of P. koraiensis. This study suggests that the traditional concept of Y‐shaped white stomatal bands in C. obtusa should be revised to describe the arrangement of the aggregated waxy stomata that occur in rows.     

A simple and efficient method for objective discrimination between pseudostratified,metaplastic and dysplastic nasal epithelium

Morphologic features may sometimes be interpreted differently by different pathologists. By introducing objective methods in the histological evaluation the subjectivity of the judgement is reduced and a higher degree of consistency achieved. In the present study three linear parameters of nasal epithelial basal cells: the largest transversal nuclear diameter, the sum of the longitudinal and transversal nucleolar axes and the basal cell width, i.e. the width of the attachment face to the basement membrane, were classified by means of three-class rulers. From this classification weighted indices were made which were then given equal weight and transformed, so that when added up their sum, the joint index (Q) to the largest possible extent was consistent with pseudostratified/stratified cuboidal, stratified squamous and dysplastic epithelium when Q ≤ 1, 1 < Q ≤ 2 and Q > 2 respectively. Tested prospectively we found 91% agreement between the morphometric and histological classification. The method presented is extremely simple and rapid to perform, and appears to have diagnostic potential in distinguishing various types of epithelial alterations of the nasal mucosa. The method may also be useful in approaching similar problems in other epithelia.     

Scanning electron microscopy of the tongue papillae in the pig (Sus scrofa)

A clear understanding of the role of different papillae in the pig may permit the development of a more palatable diet, thus, better utilizing anatomical structures and morphological characteristics to enhance health and productivity in this domestic species. The study used ten healthy market-size, mixed-breed pigs to properly describe the surface structure of the tongue papillae using standard scanning electron microscopic techniques. Two types of sharp and blunt filiform papillae were identified. Fungiform papillae, located on the lateral side of the tongue, were circular and large with surface taste pores. The majority of oval papillae on the dorsal surface of the tongue contained multidirectional surface grooves and lacked taste pores. The foliate papillae, located principally on the caudal third and lateral part of the tongue, were comprised of four to five leaves and they had additional pseudopapillae. The two vallate papillae, found in the caudal third and dorsum of the tongue, had a well-defined vallum and pseudopapillae on their surface. At the root of the tongue, two types of conical papillae with or without apex were observed. The entire area covered by conical papillae was deeply occupied by lingual tonsils. The lingual tonsils were scattered with varying sized lymph nodules in the propria mucosa of the conical papillae deep to epithelium. Functionally, the filiform and conical papillae appeared to have a mechanical role whereas fungiform, foliate, and vallate papillae apparently perform a gustatory role in the pig. The tonsillar location at the root of the tongue provides an ideal experimental model for immunohistological and immunological studies on the lymphoid tissue in man.     

Comparison between two different methods for evaluating rumen papillae measures related to different diets

Although the Geographical Information System (GIS), which integrates computerized drawing computer assisted design (CAD) and relational databases (data base management system (DBMS)), is best known for applications in geographical and planning cartography, it can also use many kinds of information concerning the territory. A multidisciplinary project was initiated since 5 years a multidisciplinary study was initiated to use GIS to integrate environmental and ecological data with findings on animal health, ethology, and anatomy. This study is chiefly aimed at comparing two different methods for measuring the absorptive surface of rumen papillae. To this scope, 21 female sheep (Ovis aries) on different alimentary regimes (e.g., milk and forage mixed diet, early herbaceous diet, dry hay diet, and fresh hay diet at the maximum of pasture flowering and at the maximum of pasture dryness) were used; after slaughtering, 20 papillae were randomly removed from each sample collected from four indicator regions of rumen wall, placed near a metric reference and digitally photographed. The images were developed with the ArcGIS? software to calculate the area of rumen papillae by means of GIS and to measure their mid‐level width and length to calculate the papillae area as previously performed with a different method. Spatial measurements were analyzed using univariate and multivariate methods. This work demonstrates that the GIS methodology can be efficiently used for measuring the absorptive surface of rumen papillae. In addition, GIS demonstrated to be a rapid, precise, and objective tool when compared with previously used method. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.     

Comparative structural morphometry and elemental composition of three marine sponges from Western Coast of India

Three marine sponges Halichondria glabrata, Cliono lobata, and Spirastrella pachyspira from the western coastal region of India were compared for their morphometry, biochemical, and elemental composition. One‐way analysis of variance was applied for spicule morphometry results. Length, width, and length:width ratio were calculated independently. The ratio of length:width varied from 35 to 42 among the grown samples, which remained in the range of 10–22 in young sample at the beginning of studies. However, no significant change was observed in spicule width compared to length. Elemental compositions of marine sponges were determined by field emission gun‐scanning electron microscope. Scanning electron microscopy data revealed that the spicules of all the three sponges were mostly composed of O (47–56%) and Si (30–40%), whereas Al (14.33%) was only detected in the spicules of C. lobata. Apart from these, K, Ni, Ca, Fe, Mg, Na, and S were additionally detected in all the three samples. Presence of heavy metals in the sponges was analyzed by inductively coupled plasma‐atomic emission spectroscopy. Results showed that iron was present in a large amount in samples, followed by zinc, lead, and copper. Microsc. Res. Tech. 77:296–304, 2014. © 2014 Wiley Periodicals, Inc.     

Junctional complex revisited by high-resolution scanning electron microscopy

The present study correlates the ultrastructural morphology of junctional complexes as revealed by transmission electron microscopy (TEM) with that observed by high-resolution scanning electron microscopy (HRSEM), thanks to a new modification of the osmium tetroxide maceration technique. The removal of all cytoplasmic organelles by this technique allows the inspection of the inner side of the plasmalemma. With this treatment, a continuous band of tightly packed particles is observed at the most apical portion of lateral membranes. Just below this band, irregular clusters of apparently identical particles are placed all around the cellular contour. The topographical correspondence among these clusters and spot desmosomes seen by TEM identifies them as desmosomes. The continuous band seems to represent the combination of both zonulae, occludens and adherens. Regarding the nature of the particles, we suppose that they probably consist of peripheral membrane proteins clustered at the cytoplasmic surface of intercellular junctions and involved in the linkage between cytoskeleton and plasmalemma.     

Scanning electron microscopic and histological studies of the buccal cavity of a phytoplanktivorous small freshwater fish,Amblypharyngodon mola

The electron microscopic and histological studies of the buccal‐cavity of herbivorous fish Mola (Amblypharyngodon mola) were performed. The studies revealed that the architectures of the buccal cavity of A. mola support the herbivory nature of the fish. Both the upper and lower jaws of the fish are rich in mucus glands, unculi, and microridges. The presence of papillae like taste buds in the lower jaw of A. mola indicates the mechanosensory role of the lower jaw during gustation. These features directly support a gustatory feeding behavior associated with filter feeding in this small freshwater fish.     

Morphological investigations on the lips and cheeks of the goat (Capra hircus): A scanning electron and light microscopic study

The current study was done to provide comprehensive information on the anatomical features of the lips and cheeks of the goat by gross examination and morphometric analysis in addition to scanning electron microscope (SEM). Samples from 12 normal healthy adult goat's heads of both sexes were collected directly after slaughtering. The lips and cheeks were dissected, and specimens were collected for both light and SEM. The lips of goat were soft and mobile. The free border of both lips was characterized rostrally by the presence of labial projections. The number, size, and arrangement of labial projections differed in the upper and lower lips. On the other hand, the buccal papillae were arranged into 6–8 longitudinal rows parallel to the cheek teeth. The length of these papillae decreased caudally while they were absent on the most caudal part of the cheek. Presence of several types and shapes of labial projections and papillae, and buccal papillae suggest a high degree of mechanical adaptation of the lips and cheeks of the goat. This study provides baseline data for clinical studies. This study is the first report to shed light on the morphology of the lips and cheeks of the goat by gross and scanning electron microscopy.