Differentiation of the tegument and associated structures in Diphyllobothrium dendriticum Nitsch (1824) (Cestoda: Pseudophyllidea). An electron microscopical study

Grammeltvedt Anne-François 1973. Differentiation of the tegument and associated structures in Diphyllobothrium dendriticum Nitsch 1824 (Cestoda : Pseudophyllidea). An electron microscopical study. International Journal for Parasitology3: 321–327. The differentiation of the tegument and associated structures of the coracidium, procercoid, plerocercoid and adult is described. The embryophore is composed of four zones and is covered by a fibrous layer resembling a glycocalyx. The oncospheral plasma membrane is extensively folded. A typical cestode tegument, with a distal and perinuclear cytoplasm, is probably already existing in the coracidium. The formation of the microvilli starts after about three days in the copepod host. In young procercoids ribosomes and Golgi complexes were observed in the distal cytoplasm. These organelles disappear at later stages. The infective procercoid has a typical tegument. The microvilli are shaped like a thorn compressed from the sides. They have an electron dense tip and a less dense base in which microfilaments are seen. Bodies, called disc-shaped and lamellated bodies, are described. The microvilli of the plerocercoid are characterized by a great variation in shape. The villi are bounded by two unit membranes. The lamellated bodies are especially well developed. The adult microvilli are uniform in shape. The lamellated bodies are few in young adults and disappear in mature worms.     

A study of the microtriches of adult Echinococcus granulosus by scanning electron microscopy

Thompson R. C. A., Houghton A. and Zaman V. 1982. A study of the microtriches of adult Echinococcus granulosus by scanning electron microscopy. International Journal for Parasitology12: 579–583. The microtriches in different regions of the scolex and strobila of sexually mature Echinococcus granulosus were examined using the scanning electron microscope. E. granulosus was found to possess two morphologically distinct types of microthrix. A cylindrical, slender filamentous type of microthrix which appeared to be flexible was restricted to the scolex. On the strobila, the only type of microthrix observed was a flattened blade-like form which appeared to be rigid for most of its length. The occurrence of two distinct types of microthrix in separate regions of the body of E. granulosus suggests that they may be involved in different functional activities.     

An electron microscope study of the tegument of Hunterella nodulosa Mackiewicz and McCrae, 1962 (cestoidea: Caryophyllidea)

Ultrastructural observations using transmission and scanning electron microscopy reveal the tegument to be basically similar to that of other cestodes. The syncytial distal cytoplasm is devoid of organelles except for rod-shaped bodies, believed to be secretory vesicles, and lamellated bodies which probably contribute the raw material for new microtriches. There is evidence that these vesicles originate from the Golgi found in the sub-cuticular cells.Three types of microtriches are described: typical ones with well-developed spines, ones with short filaments instead of spines, and ones with no spines. Microtriches with spines are found only on the anterior part of the worm and may serve to anchor the worm. Microtriches on the posterior have no spines and are believed to be primarily involved in the absorption of nutrients. Between these two regions there is a transitional zone where all three types of microtriches can be found. In general the microtriches are quite uniformly distributed throughout the surface of the worm. The presence of cestodarian-like microtriches raises interesting evolutionary questions.Histochemical tests localized acid and alkaline phosphatase activity on various parts of the tegument, as well as on host intestine, while acian blue tests showed that acid mucopolysaccharide levels correspond with the concentration of the tegument vesicles.     

Identification of telocytes in the lamina propria of rat duodenum: transmission electron microscopy

Recently the new term 'telocytes' has been proposed for cells formerly known as interstitial Cajal-like cells. In fact, telocytes are not really Cajal-like cells, they being different from all other interstitial cells by the presence of telopodes, which are cell-body prolongations, very thin, extremely long with a moniliform aspect. The identification of these cells is based on ultrastructural criteria. The presence of telocytes in others organs was previously documented. We reported for the first time, an ultrastructural study of telocytes in the lamina propria of rat duodenum. Our findings show that typical telocytes are present in the rat duodenum. Telocytes are located in the lamina propria, immediately below mucosal crypts. Telopodes frequently establish close spatial relationships with immune cells, blood vessels and nerve endings. On the basis of their distribution and morphology, we suggest that these cells may be involved in immune response and in our opinion, it may be possible that different locations of telocytes could be associated with different roles.     

Effects of millimeter-wave radiation on monolayer cell cultures. II. Scanning and transmission electron microscopy

Both thermal and athermal effects of millimeter-wave radiation on BHK-21/C13 cells were sought using scanning and transmission electron microscopy in conjunction with an in vitro technique that allows direct exposure of monolayer cultures to high average power densities. Culture dishes were irradiated by placing them on the open end of an E- or U-band wave guide. This technique exposes different regions of the cell monolayer lying along the longer axis of the wave guide aperture to varying power densities ranging from zero at each edge to twice the average power density at the center. Cell ultrastructure was unaffected by microwave radiation for 1 hour (41.8 or 74.0 GHz, average power densitites = 320 or 450 mW/cm², respectively) with or without cooling by rapid recirculation of the culture medium. Temperature in recirculated cultures was held at 37.2 °C, and that in noncooled cultures never exceeded 42 °C during irradiation at either power density. In contrast, cell morphology was affected by microwave exposure whenever irradiation conditions were altered so that the temperature of the monolayer reached or exceeded 44.5 °C. Ultrastructural alterations included breakage of cell processes, progressive detachment of cells from the substrate, increased clumping of heterochromatin in the nuclei, and the appearance of large empty vesicles in the cytoplasm. Such morphological changes resulted from either application of higher average power densities or irradiation at the power densities described above at a higher ambient temperature (>38.5°C).     

Echinococcus multilocularis in the cotton rat: the in vitro protoscolicidal activity of peritoneal cells.

Protoscolices of Echinococcus multilocularis were incubated in vitro with peritoneal cells and with sera from normal and infected cotton rats. The cells from rats bearing massive subcutaneous and intraperitoneal hydatid cysts rapidly killed protoscolices; normal cells and cells from lightly infected animals did not. Only sera from rats bearing large, subcutaneous cysts displayed significant protoscolicidal activity. These data suggest that the phenomenon whereby large, established cysts of E. multilocularis effectively suppress the establishment, growth, and metastasis of distant foci of infection has an immunological component.     

Low-temperature electron microscopy for the study of polysaccharide ultrastructures in hydrogels. I. Theoretical and technical considerations

The high-pressure freezing (HPF) technique was applied to the cryo-immobilization of alginate gels and the quality of the freezing analyzed on a TEM by comparison of the segregation pattern of samples of decreasing thickness. Dynamic simulations of heat transfer within an idealized slab of pure water surrounded by two walls of aluminium were performed to illustrate the effect of the heat-transfer coefficient by convection on the cooling rate of the sample. Heat-transfer coefficients in liquid nitrogen and liquid propane at ambient pressure were measured using a carefully characterized thermocouple and the values incorporated as parameters in heat-transfer simulations to compare the efficiency of the plunge-freezing technique with the high-pressure freezing technique. Values of the heat-transfer coefficient in liquid nitrogen and liquid propane, calculated between 273 K and 173 K were 670 and 18420 W/m(2)/K, respectively. Based on TEM observations and the results of heat-transfer simulations, the HPF technique was adapted to the cryo-fixation of 50-microm-thick alginate gels. The occurrence of artifacts was rejected because no differences were observed in the pattern of cryo-fixed and freeze-substituted samples of various thickness, with and without ethanol as cryo-protectant. A sample thickness of 50 microm was found to ensure an adequate preservation of structures as small as a few nanometers, as verified by TEM and SEM observations. Finally, DSC measurements on alginate solutions and alginate beads revealed that under the experimental conditions (0-3%), alginate cannot be considered to be an efficient cryo-protectant.     

Gills of the medaka (Oryzias latipes): A scanning electron microscopy study

The general morphology and surface ultrastructure of the gills of adult and larvae medaka (Oryzias latipes) were studied in freshwater and seawater using scanning electron microscopy. The gills of all examined fish were structurally similar to those of other teleosts and consisted of four pairs of arches supporting (i) filaments bearing lamellae and (ii) rakers containing taste buds. Three cell types, specifically pavement cells, mitochondria‐rich cells (MRCs), and mucous cells, constituted the surface layer of the gill epithelium. Several distinctive characteristics of medaka gills were noted, including the presence of regularly distributed outgrowth on the lamellae, enlarged filament tips, the absence of microridges in most pavement cells in the filament and lamellae and the presence of MRCs in the arch at the filament base. A rapid mode of development was recorded in the gills of larval fish. At hatching, the larvae already had four arches with rudimentary filaments, rakers, and taste buds. The rudimentary lamellae appeared within 2 days after hatching. These results suggest the early involvement of larval gills in respiratory and osmoregulation activities. The responses of the macrostructures and microstructures of gills to seawater acclimation were similar in larvae and adult fish and included modification of the apical surface of MRCs, confirming the importance of these cells in osmoregulation. The potential roles of these peculiarities of the macrostructures and microstructures of medaka gills in the major functions of this organ, such as respiration and osmoregulation, are discussed.     

Echinococcus granulosus: study of the in vitro complement activation by protoscoleces by measuring the electric potential difference across the tegumental membrane.

Complement activation by protoscoleces of Echinococcus granulosus was studied by analyzing the damage to their tegumental membrane produced by incubation in both normal and hydatid human sera. The state of the apical tegumental membrane was evaluated by measuring the electric potential difference with microelectrodes. Protoscoleces incubated in Ringer-Hepes or in heat-decomplemented normal human serum in the presence or absence of specific antibodies did not show significant variations in the electric potential difference throughout the experiment (P > 0.4 in all cases) and their mean values were -46 +/- 3, -43 +/- 4, and -56 +/- 5 mV, respectively. In contrast the potential difference of protoscoleces incubated in 1:2 diluted normal human serum showed a significant variation (P < 0.001), reaching -10 +/- 6 mV after 30 min, and the median depolarization time was estimated to be 21 +/- 3 min. The capacity of normal human serum to depolarize the tegumental membrane of protoscoleces was abolished by treatment at 50 degrees C during 20 min or by 10-fold dilution. In addition, protoscoleces incubated in 1:10 diluted hydatid human serum plus 1:10 diluted normal human serum or Factor B-inactivated normal human serum showed a significantly faster depolarization (0.01 < P < 0.02 and P < 0.001, respectively): the potential difference reached -13 +/- 5 mV after 15 min and the median depolarization times were 9 +/- 5 and 5 +/- 3 min, respectively. Our results suggest that following the time course of the potential difference is a useful tool for studying complement activation in the host-parasite interface and they show that the tegumental membrane of protoscoleces can activate the alternative pathway of human complement.     

Antifungal activity of allylamines on Epidermophyton floccosum: scanning electron microscopy study

The action of allylamine antifungal agents on Epidermophyton floccosum was studied using scanning electron microscopy. After 7 days of culture on Sabouraud dextrose agar, Epidermophyton floccosum samples were brought in contact with concentrations of 0.2 and 2 g ml^–1 and 0.01 and 0.1 g ml^–1 of naftifine and terbinafine, respectively. Lesions observed after 24 h, 3 and 7 days of contact were mainly on the structure and rigidity of the mycelial and macroconidial wall. They were characterized by hyphal ballooning and twisting and by apical bulbous bulges. Deterioration of macroconidia was characterized by wall exfoliation. The intensity of the deterioration depended on the dose and only slightly on the length of time that the sample and the antifungal drug were in contact.     

Adhesion of Plasmodium gallinaceum Ookinetes to the Aedes aegypti Midgut: Sites of Parasite Attachment and Morphological Changes in the Ookinete

Plasmodium gallinaceum ookinetes adhered to Aedes aegypti midgut epithelia when purified ookinetes and isolated midguts were combined in vitro. Ookinetes preferentially bound to the microvillated luminal surface of the midgut, and they seemed to interact with three types of structures on the midgut surface. First, they adhered lo and migrated through a network-like matrix, which we have termed microvilli-associated network, that covers the surface of the microvilli. This network forms on the luminal midgut surface in response to blood or protein meals. Second, the ookinetes bound directly to the microvilli on the surface of the midgut and were occasionally found immersed in the thick microvillar layer. Third, the ookinetes associated with accumulations of vesicular structures found interspersed between the microvillated cells of the midgut. The origin of these vesicular structures is unknown, but they correlated with the surface of midgut cells invaded by ookinetes as observed by TEM. After binding to the midgut. ookinetes underwent extensive morphological changes: they frequently developed one or more annular constrictions, and their surface roughened considerably, suggesting that midgut components remain bound to the parasite surface. Our observations suggest that, in a natural infection, the ookinete interacts in a sequential manner with specific components of the midgut surface. Initial binding to the midgut surface may activate the ookinete and cause morphological changes in preparation for invasion of the midgut cells.     

粉尘螨生殖系统超微结构的透射电镜观察

本研究主要采用透射电镜观察粉尘螨Dermatophagoides farinae (Hughes)生殖系统超微结构。粉尘螨雄性生殖系统是由精巢、 输 精管、 附腺、 射精管、 交配器官及附属交配器官组成。精巢内可同时有精子发育各阶段的细胞。精子无核膜、 核染色质聚集成束、 线 粒体缺乏典型的嵴、 胞质内有平行排列的电子致密薄片等为其特征性结构。雌性生殖系统由交合囊、 交合囊管、 储精囊、 囊导管、 卵 巢、 输卵管、 子宫及产卵管构成。卵巢内可见含多个细胞核的中央细胞, 其周为卵母细胞等生殖细胞。该研究丰富了对粉尘螨生殖系统 结构的认识。     

Morphological specializations of the buccal cavity in relation to the food and feeding habit of a carp Cirrhinus mrigala: A scanning electron microscopic investigation

The buccal cavity of an herbivorous fish, Cirrhinus mrigala, was investigated by scanning electron microscopy to determine its surface ultrastructure. The buccal cavity shows significant adaptive modifications in relation to food and feeding ecology of the fish. The buccal cavity of the fish is of modest size and limited capacity, which is considered an adaptation with respect to the small‐sized food items primarily consumed by the fish that could be accommodated in a small space. Modification of surface epithelial cells, on the upper jaw, into characteristic structures—the unculi—is considered an adaptation to browse or scrap, to grasp food materials, e.g., algal felts, and to protect the epithelial surface against abrasions, likely to occur during their characteristic feeding behavior. Differentiation of the highly specialized lamellar organ on the anterior region of the palate could be an adaptation playing a significant role in the selection, retention, and sorting out of palatable food particles from the unpalatable items ingested by the fish. The filamentous epithelial projections and the lingulate epithelial projections on the palatal organ in the posterior region of the palate are considered to serve a critical function in final selection, handling, maneuvering, and propelling the food particles toward the esophagus. The abundance of different categories of taste buds in the buccal cavity suggests that gustation is well developed and the fish is highly responsive in the evaluation and the selection of the preferred palatable food items. The secretions of mucous cells in the buccal cavity are associated with multiple functions—particle entrapment, lubrication of the buccal epithelium and food particles to assist smooth passage of food, and to protect the epithelium from possible abrasion. These morphological characteristics ensure efficient working of the buccal cavity in the assessment of the quality and palatability of ingested food, their retention and transport toward the esophagus. Such an adaptation may be essential in conducting the function most basic to the survival of the individuals and species—feeding. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.     

Ultrastructure of Red-Sore Lesions on Largemouth Bass (Micropterus salmoides): Association of the Ciliate Epistylis sp. and the Bacterium Aeromonas hydrophila*

SYNOPSIS. Epizootic outbreaks of red-sore disease in several reservoirs in the southeastern United States have been reported to cause heavy mortality among several species of fish having sport and commercial value. The etiologic agent is said to be the peritrich ciliate Epistylis sp.; secondary infection by the gram-negative bacterium Aeromonas hydrophila produces hemorrhagic septicemia which results in death. However, in recent studies on the largemouth bass Micropterus salmoides, Epistylis sp. could be isolated from only 35% of 114 lesions from 114 fish, while A. hydrophila was found in 96% of the same lesions. Transmission and scanning electron microscopy of lesions associated with red-sore disease indicate that neither the stalk nor the attachment structure of Epistylis sp. have organelles capable of producing lytic enzymes. Since other investigators have shown that A. hydrophila produces strong lytic toxins, and in absence of evidence to the contrary, it is concluded that Epistylis sp. is a benign ectocommensal and that A. hydrophila is the primary etiologic agent of red-sore disease.     

Organization of the olfactory system of the Indian major carp <Emphasis Type="Italic">Labeo rohita</Emphasis> (Ham.): A scanning and transmission electron microscopy study

Catla catla, Labeo rohita, and Cirrhinus mrigala represent important alimentary fish in India. Their reproduction/breeding depends on seasons. Fish perceive external factors-stimuli and chemical signals through the olfactory system that plays the key role in central regulation of reproduction. However, no electron microscopy data are available on organization of olfactory components of these fish. We studied organization of the olfactory organ in male L. rohita using scanning (SEM) and transmission electron microscopy (TEM). This organ consists of olfactory epithelium, a short nerve, and olfactory bulb. The olfactory organ is ovoid in shape and consists of about 47–52 lamellae in adults and about 14–20 lamellae in fingerlings. These lamellae originate from the midline raphe. By SEM, microvillar sensory and ciliated non-sensory cells were observed in the lamellae. TEM revealed microvillar receptor cell with rough endoplasmic reticulum and Golgi apparatus towards apical end. Basal cells were present at the base of receptor cell, supporting cells were located adjacent to the olfactory receptor neurons, while epithelial cells—in the nonsensory part of olfactory epithelium. Mast, blastema, and macrophage cells were also found at the basement membrane. This work is the first publication on ultrastructural organization of the olfactory system of the Indian major carp, which provides information about morphological and ultrastructural organization of the olfactory system and opens new avenues for further investigation of chemical neuroanatomy, sensory signal processing, and neural regulation of reproduction in the Indian major carp.     

Observations on the coiled posture of trichostrongyle infective larvae using a freeze-substitution method and scanning electron microscopy

Wharton D. A. 1982. Observations on the coiled posture of trichostrongyle infective larvae using a freeze-substitution method and scanning electron microscopy. International Journal for Parasitology12: 335–343. A method for the preparation of the infective larvae of Trichostrongylus colubriformis for scanning electron microscopy is described. This involves freeze-substitution with methanol followed by critical-point drying. The method resulted in good preservation and the retention of the coiled posture. The morphology of ensheathed and exsheathed larvae is described and the mechanisms of coil formation discussed. Coiling was also observed in the infective larvae of nine other trichostrongyle species.     

Effect of selenium compounds on the damage induced by oxysterol on rat arterial walls

Wistar rats were fed Se-deficient (0.017±0.002 mg Se/kg) and Seadequate (0.32±0.045 Se mg/kg) diets for 12 mo and then were given 5 mg/kg of cholestane-3β,tα,6β-triol (3-triol), intravenously. Se compounds (Na₂SeO₃ and ebselen) were supplemented in different doses and times to the Se-deficient rats. Twenty-four hours after 3-triol infusion, the changes in ultrastructures of rat aorta were examined by scanning electron micrography (SEM) and transmission electron micrography (TEM). SEM examinations showed that 3-triol induced diffused injuries on arterial endothelial urfaces of long-term Se-deficient rat, and a large number of holes or craterlike defects were observed. TEM examinations further showed that 3-triol induced swelling, necrosis, and shedding of endothelial cells, which resulted in the destruction of endothelial integrity. Mean-while, smooth muscle cells proliferated and migrated toward intimae; the breakage of internal elastic lamina benefited the migration of smooth muscle cells. Supplemented with Na₂SeO₃ (40 μg/kg, 10 d per continuum) and ebselen (20 mg/kg), respectively, exhibited significant protection from damages induced by 3-triol. It seems that protecting mechanisms were different between Na₂SeO₃ and ebselen. The present investigation gave visual evidence that both injuries induced by cholesterol oxides and the Se nutritional status contributed to the development of atherosclerosis.     

A scanning electron microscope study of a microsporidian with a pansporoblast: Thelohania maenadis

Air drying is not adequate for the preservation of the pansporoblastic membrane of Thelohania maenadis (Protozoa, Microsporidia), a parasite of the crabs Carcinus mediterraneus and Carcinus maenas. Freeze-drying and critical point drying preserve the pansporoblast membrane and reveal that isolated spores of the microsporidian are covered with a thick hairy coat. This coat originates as secretory product within the pansporoblast cavity.     

Transthyretin fibrillogenesis entails the assembly of monomers: a molecular model for in vitro assembled transthyretin amyloid-like fibrils

Extracellular accumulation of transthyretin (TTR) variants in the form of fibrillar amyloid deposits is the pathological hallmark of familial amyloidotic polyneuropathy (FAP). The TTR Leu55Pro variant occurs in the most aggressive forms of this disease. Inhibition of TTR wild-type (WT) and particularly TTR Leu55Pro fibril formation is of interest as a potential therapeutic strategy and requires a thorough understanding of the fibril assembly mechanism. To this end, we report on the in vitro assembly properties as observed by transmission electron microscopy (TEM), atomic force microscopy (AFM) and quantitative scanning transmission electron microscopy (STEM) for both TTR WT fibrils produced by acidification, and TTR Leu55Pro fibrils assembled at physiological pH. The morphological features and dimensions of TTR WT and TTR Leu55Pro fibrils were similar, with up to 300 nm long, 8 nm wide fibrils being the most prominent species in both cases. Other species were evident; 4-5 nm wide fibrils, 9-10 nm wide fibrils and oligomers of various sizes. STEM mass-per-length (MPL) measurements revealed discrete fibril types with masses of 9.5 and 14.0(+/-1.4) KDa/nm for TTR WT fibrils and 13.7, 18.5 and 23.2(+/-1.5) kDa/nm for TTR Leu55Pro fibrils. These MPL values are consistent with a model in which fibrillar TTR structures are composed of two, three, four or five elementary protofilaments, with each protofilament being a vertical stack of structurally modified TTR monomers assembled with the 2.9 nm axial monomer-monomer spacing indicated by X-ray fibre diffraction data. Ex vivo TTR amyloid fibrils were examined. From their morphological appearance compared to these, the in vitro assembled TTR WT and Leu55Pro fibrils examined may represent immature fibrillar species. The in vitro system operating at physiological pH for TTR Leu55Pro and the model presented for the molecular arrangement of TTR monomers within fibrils may, therefore, describe early fibril assembly events in vivo.