Ultrastructural description of an unidentified apicomplexan oocyst containing bacteria-like hyperparasites in the gill of Crassostrea rizophorae

Oocysts of an unidentified coccidian are reported in this study to parasitize the gills of the oyster Crassostrea rizophorae (Mollusca, Bivalvia) collected near the city of Recife (Itamaracá Island, 07 degrees 38' 00" S, 34 degrees 48' 06" W), Brazil. Oocysts appeared as light and dense forms, both containing rod-shaped, bacteria-like hyperparasites (BL). Both light and dense oocysts were spherical, 4.3 to 4.7 pm in diameter, but denser oocysts had irregular contours. Both forms consisted of a thick dense wall (approximately 165 nm thick) consisting of 3 layers. The outermost, a dense and irregular layer about 25 nm thick, possessed numerous bead-like structures and some slender conical projections (up to 1.5 microm long). The inner layer of the wall was formed by a dense and homogenous layer about 125 nm thick. Between these 2 layers, a thin light layer about 12 nm thick was present. Uninucleated sporocysts occupied the internal space of the oocyst and contained some rod-shaped BL and mitochondria surrounded by numerous ribosome-like particles. The dense forms of the oocysts showed the same structures described in the lighter forms and appeared to be the final maturation form of the oocysts. Free sporozoites were occasionally observed among oocysts.     

呼肠孤病毒与SARS相关的形态学依据

SARS患者病理尸检肺组织样品分离病毒出现细胞病变的Hep2 培养细胞,按常规制作超薄切片,透射电镜下观察。电镜下,检出在感染细胞内复制、组装的呼肠孤病毒及其包涵体。病毒粒子衣壳立体对称、无包膜、直径在60~80nm。成熟病毒粒子核心致密常排列呈晶格状,不成熟病毒粒子核心空亮。数目不等的上述两种病毒粒子、长短不等的微管样结构和病毒浆常在核旁胞质内组成大小不等、无定形的病毒包涵体。此发现进一步提供了呼肠孤病毒感染有可能与SARS相关的形态学依据。     

Surface coat transformation and capsule formation by Leuconostoc mesenteroides NCDO 523 in the presence of sucrose

When Leuconostoc mesenteroides NCDO 523 was grown in MRS browth, electron microscopy of cells fixed in the presence of ruthenium red showed that the cell wall was covered with a thin layer of filamentous material. When MRS-grown cells were resuspended in the same medium supplemented with 3.6% sucrose, this surface coat doubled in thickness and a number of radial thickenings appeared within it. After 3 h the filamentous component of the surface coat had disappeared leaving only the radial projections. The progressive accumulation of polymer to produce a capsule visible by light microscopy was observed in only about 20% of the population. In this minority of cells, a dense globular dextran composed of fibrillar and particulate elements was always produced in the initial stages of synthesis. After 18 h, the dextran capsule was generally composed of an inner globular and outer fibrillar layer. It appeared that the outer layer was derived from the globular dextran of the capsule by a process of dispersion.     

Diffusion of macromolecules and virus-like particles in human cervical mucus

To determine whether or not large macromolecules and viruses can diffuse through mucus, we observed the motion of proteins, microspheres, and viruses in fresh samples of human cervical mucus using fluorescent recovery after photobleaching and multiple image photography. Two capsid virus-like particles, human papilloma virus (55 nm, approximately 20,000 kDa) and Norwalk virus (38 nm, approximately 10,000 kDa), as well as most of the globular proteins tested (15-650 kDa) diffused as rapidly in mucus as in saline. Electron microscopy of cervical mucus confirmed that the mesh spacing between mucin fibers is large enough (20-200 nm) for small viruses to diffuse essentially unhindered through mucus. In contrast, herpes simplex virus (180 nm) colocalized with strands of thick mucus, suggesting that herpes simplex virus, unlike the capsid virus particles, makes low-affinity bonds with mucins. Polystyrene microspheres (59-1000 nm) bound more tightly to mucins, bundling them into thick cables. Although immunoglobulins are too small to be slowed by the mesh spacing between mucins, diffusion by IgM was slowed by mucus. Diffusion by IgM-Fc(5 mu), the Fc pentamer core of an IgM with all 10 Fab moieties removed, was comparably slowed by mucus. This suggests that the Fc moieties of antibodies make low-affinity bonds with mucins.     

The fine structure of the developing oocyst of Pterospora floridiensis (Apicomplexa,Urosporidae)

Developing oocysts of the gregarine Pterospora floridiensis Landers 2001 were examined by transmission electron microscopy. Each oocyst had an outer capsule and an inner capsule that contained 8 sporozoites. In early stages of development the inner capsular wall was separated from the developing sporozoites and residual mass, and was not appressed to the sporozoites. Early stage sporozoites were connected to a residual mass and were filled with endoplasmic reticulum, golgi and numerous developing secretory vesicles. In late stages of oocyst and sporozoite development, the inner capsular wall was closely appressed to the sporozoite surface. The inner capsular wall was ～60-100 nm thick and the outer capsular wall was ～160-320 nm thick. There were no extensions on the outer wall for which the genus was named. Late stage sporozoites had no residual mass connection, were more electron dense, and contained three distinct types of dense secretory structures: 1) small oval/spherical dense vesicles, 2) large (350-400 nm) vesicles near the anterior end, and 3) elongated dense tubular bodies that converged at the apex. Few ultrastructural reports exist of developing gregarine oocysts and sporozoites, and as more studies are completed these morphological characteristics may be important in interpreting molecular phylogenetic analyses.     

Spermatogenesis and sperm structure of Acerella muscorum, (Ionescu, 1930) (Hexapoda, Protura)

The general organization of the male genital system, the spermatogenesis and the sperm structure of the proturan Acerella muscorum have been described. At the apex of testis apical huge cells are present; their cytoplasm contains a conventional centriole, a large amount of dense material and several less electron-dense masses surrounded by mitochondria. Spermatocytes have normal centrioles and are interconnected by cytoplasmic bridges. Such bridges seem to be absent between spermatid cells and justify the lack of synchronization of cell maturation. Spermatids are almost globular cells with a spheroidal nucleus and a large mass of dense material corresponding to the centriole adjunct. Within this mass a centriole is preserved. Mitochondria of normal structure are located between the nucleus and the plasma membrane. The spermatids are surrounded by a thick membrane. No flagellar structure is formed. Sperm have a compact spheroidal nucleus, a large cap of centriole adjunct material within which a centriole is still visible. A layer of mitochondria is located over the nucleus. The cytoplasm is reduced in comparison to spermatids; many dense bodies are interspersed with sperm in the testicular lumen. The sperm are small, immotile cells of about 2.5-3 μm in diameter.     

Die Feinstruktur der lamellären Einschlußkörper in den Zellkernen des Nektariums von Catalpa bungei

Summary In the nectary of Catalpa bungei the gland cells contain nuclear inclusions consisting of stacks of lamellae 12.7 nm thick. Each lamella is composed of globular particles with a diameter of approximately 12.7 nm. The particles are arranged in a monolayer, revealing a regular square pattern in face view. In adjacent lamellae the globular subunits are almost exactly superimposed; each of them is probably built up of smaller subunits having a diameter of 0.4 nm.

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Herrn Prof. Dr. Horst Drawert nachträglich zum 60. Geburtstag gewidmet     

Chromatin structure within sites of DNA replication

Conformational changes in chromatin structure are nowadays the object of intensive research due to its importance for proper regulation of intranuclear processes. The fine structure of chromatin within the DNA replication sites was studied in in situ fixed cells and cells permebilized by low ionic strength solutions in the presence of divalent cations. The latter method provides visualization of higher level chromatin structures such as globular chromomeres and chromonema fibres. Nascent DNA was detected immunochemically using anti-BrdU antibodies on the surface of ultrathin sections prepared from Epon-embedded material. It was shown that newly replicated DNA preferentially localized within the zones filled with globular and fibrillar elements with characteristic diameter of 30 nm, and not in chromonema fibres, while after replication had been completed DNA became embedded into as thick as 60-80 nm chromonema elements. The results obtained are discussed in the context of conception of hierarchical folding of chromatin fibers.     

An Ultrastructural and Cytochemical Study of the Cell Surface of a Suctorian Ciliate1

The surfaces of the main cell body, tentacle shaft, and knob of Discophrya collini, a freshwater suctorian ciliate, were characterized using various cytochemical techniques. Cells prepared for conventional transmission electron microscopy exhibited a 50–60 nm thick fuzzy layer over the cell body surface; this layer was absent from the tentacle knob. A thick (240 nm), two-layered surface coat surrounding the main cell body was stained with ruthenium red. This heavy coat was absent from the surface of the knob where a thin, dense, ruthenium red-positive layer and projecting filaments were present. Freeze-etched material revealed a “particle region” (150–250 nm in thickness) closely associated with the outer cell surface of the suctorian. Fixed specimens were treated with four different lectins and analyzed with electron microscopy in order to obtain information about the carbohydrate composition of the outer surface of D. collini. Concanavalin A bound to the surface of the cell body and tentacle shaft as a dense, particulate layer (80 nm thick) but thinned to 13–16 nm over the surface of the knob. Wheat germ agglutinin-treated cells also displayed a heavy, electron-dense layer (128 nm thick) that surrounded the main cell body and tentacle shaft, but only scattered patches of bound wheat germ agglutinin were observed on the surface of the knob. Discophrya treated with Helix agglutinin or peanut agglutinin appeared similar to control cells. Suctorians were treated with lectins in vivo in an attempt to inhibit capture and ingestion of their prey, Tetrahymena pyriformis, by masking prey receptor sites on the knob. Concanavalin A and, to a lesser degree, wheat germ agglutinin, successfully inhibited attachment of the prey organism. Helix agglutinin and peanut agglutinin had little effect on prey capture.     

Morphogenic events during indirect somatic embryogenesis in coffee “Catimor”

Summary The main goal of this research was to identify and describe the morphological and histological events during coffee somatic embryogenesis. Leaf sections of coffee Catimor (Coffea arabica CV. Red Caturra X hybrid of Timor) were cultivated in vitro on solid medium containing 2,4-dichlorophenoxyacetic acid and benzyladenine. After 4 months, the calli produced were transferred to a medium containing naphthalene acetic acid. During the process of somatic embryogenesis, calli were sampled for histological observation. After four days of culture, the expiant produced a callus in the cut edges, where cell division occurred in the spongy parenchyma and in the perivascular parenchyma. After two months of culture, the first sign of organization within the growing callus was evidenced by the formation of densely stained cell groups appearing physically isolated, surrounded by thick cell walls. Two months later, proembryogenic clumps were formed by groups of dividing cells, unconnected to the callus. These cells were small, relatively isodiametric, with a dense cytoplasm, large nucleus, prominent nucleoli and thick cell walls. Afterwards, embryogenic calli formed somatic embryos going through the typical stages of development: globular, heart, and torpedo shapes. Histological observations revealed that the somatic embryos originated from a single cell, with dense cytoplasm, prominent nucleus and with signs of isolation evidenced by the presence of a thick cell wall.Abbreviations BAP 6-benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA naphthalene acetic acid - SEM scanning electron microscopy     

Electron microscopic study of troponin

Troponin and its components or fragments were observed in an electron microscope by the use of the rotary shadowing technique. In freshly prepared troponin with low viscosity, globular particles were mainly observed. The size of the long axis of the particles was 13.2 +/- 1.3 nm and the size perpendicular to the long axis was 9.5 +/- 1.2 nm. The mean axial ratio was 1.4 +/- 0.3. Most of the particles observed in a stored troponin preparation, having a higher viscosity than that of fresh troponin, had a globular head with a thin tail, with the total length of 25.4 +/- 1.4 nm (head-tail type particles). The axial size of the globular portion was 8.3 +/- 1.2 nm and the tail length was 17.1 +/- 1.6 nm. Observation of various particles during the transitional stages indicated that, in the globular particles, the tail region of head-tail type particle was associated along the globular head region. Troponin T was a filamentous particle with 16.9 +/- 1.5 nm length. The 26K fragment of troponin T, which was devoid of the N-terminal 45 residues from troponin T, was a filamentous particle with the length of 14.4 +/- 1.3 nm. Troponin T1, one of two chymotryptic subfragments of troponin T, was a filamentous particle of 11.6 +/- 1.4 nm length. Troponin C.T in the presence of Ca2+ was a particle with a globular head (7 nm in size) and a tail of about 17 nm length. The Fab fragment of anti-troponin T1 formed regular transverse striations along the thin filament of rabbit skeletal muscle with a 38 nm period.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ultrastructural changes in contents of membrane-coating granules after extrusion from epithelial cells of hamster cheek pouch

Summary Membrane-coating granules of the epithelium of the hamster cheek pouch displayed a lamellated internal structure consisting of alternating thick and thin electron dense bands separated by translucent bands of equal width. The repeat distance of the contents was 8.5nm±sd 0.7nm and the thickness of the major electron dense band was 3 nm±sd 0.4 nm. After exocytosis from the cells the lamellated material became organised into extensive sheets and the dimensions changed so that the repeat distance was 10.9 nm±sd 0.9 nm but the major electron dense band was virtually unchanged at 3.3 nm±sd 1.1 nm. The narrower or intermediate dense band became progressively less easy to see and was virtually invisible at the surface of the epithelium.It is suggested that the changes observed may be due to an increase in hydration of a phospholipid/protein complex in the granules by permeation of water along the intermediate dense band.     

Control of Morphogenesis in Geodermatophilus: Ultrastructural Studies

Geodermatophilus grows in two major forms, a nonmotile irregularly shaped aggregate of coccoid cells (C-form) and a motile budding rod (R-form). Morphogenesis can be controlled by an unidentified factor in Tryptose which is required for maintenance of the organism in the C-form and for differentiation of the R-form to the C-form. Morphogenetic events occur synchronously in the described system. Ultrastructural studies show that the major difference between C- and R-forms is in cells envelope structure. R-form cell walls consist of two layers, an inner transparent membranous layer (10 to 12.5 nm thick) and an outer dense diffuse layer (7.5 to 10 nm). In addition to these layers, the C-form has a thick fibrous layer (30 nm) over the dense layer. This layer appears to be a cementing substance which holds the coccoid cells together.     

Ultrastructural differences between the anterior and posterior adductors of the strawberry cockle, Fragum unedo

Adductors of Fragum unedo were observed ultrastructurally and their muscle cells were classified according to the statistically analyzed diameter of their thick myofilaments. Two types of smooth muscle cells were observed in the opaque portion of the anterior adductor: A-type cells containing thick myofilaments of about 46 nm in diameter and B-type cells having 62 nm thick myofilaments. The posterior adductor was also composed of two kinds of cells: the B-type cell, which had thick myofilaments of about 67 nm in diameter, and the C-type, containing thick myofilaments of 90 nm. Two types of oblique-striated cells were commonly recognized in the translucent portions of anterior and posterior adductors. Our observations thus indicate that the posterior adductor generally consists of cells which have thicker myofilaments than the ones of the anterior adductor.     

Vavraia lutzomyiae n. sp. (Phylum Microspora) infecting the sandfly Lutzomyia longipalpis (Psychodidae, Phlebotominae), a vector of human visceral leishmaniasis

Vavraia lutzomyiae (Microsporida; Pleistophoridae) is a new species parasitic in the tropical phlebotomine sandfly, Lutzomyia longipalpis (Diptera, Psychodidae, Phlebotominae), a major vector of Leishmania chagasi in Latin America where human visceral leishmaniasis is endemic. Infected larvae and pupae were parasitized in the abdomen, and some adults were parasitized in Malpighian tubules and midgut. The sporogonial plasmodium divided by multiple divisions into up to 64 uninucleate sporoblasts. These stages were surrounded outside the plasmalemma by a thick, amorphous dense coat and transformed into a merontogenetic sporophorous vesicle within which the sporonts developed into sporoblasts. The mature microsporidian spores were broadly ellipsoidal and measured 6.1+/-0.43 x 3.1+/-0.15 microm. The spore wall consisted of a transparent endospore (approximately 100 nm) and a thin electron dense exospore (approximately 30 nm) with the outer limit slightly undulated. Spores contained a polar filament arranged peripherally in a single layer of eight to nine wide anterior coils (approximately 125 nm diameter), and three to four narrow posterior coils (approximately 70 nm diameter). Transverse sections revealed a concentric layer organization with the internal layer surrounded by numerous (up to 25) longitudinal microfibrils. The angle of tilt of the polar filament was about 65-68 degrees.     

Description of Chytridiopsis Trichopterae N. Sp. (Microspora, Chytridiopsidae), A Microsporidian Parasite of the Caddis Fly Polycentropus Flavomaculatus (Trichoptera, Polycentropodidae), With Comments On Relationships Between the Families Chytridiopsidae and Metchnikovellidae

ABSTRACT. The microsporidium Chytridiopsis trichopterae n. sp., a parasite of the midgut epithelium of larvae of the caddis fly Polycentropus flavomaculatus found in southern Sweden, is described based on light microscopic and ultrastructural characteristics. All life cycle stages have isolated nuclei. Merogonial reproduction was not observed. the sporogony comprises two sequences: one with free spores in parasitophorous vacuoles, the other in spherical, 5.6-6.8 μm wide, sporophorous vesicles which lie in the cytoplasm. the free sporogony yields more than 20 spores per sporont. the vesicle-bound sporogony produces 8, 12 or 16 spores. the envelope of the sporophorous vesicle is about 82 nm thick and layered. the internal layer is the plasma membrane of the sporont; the surface layer is electron dense with regularly arranged translucent components. Both spore types are spherical. They have an ～ 35-nm thick spore wall, with a plasma membrane, an electron-lucent endospore, and an ～ 14-nm thick electron-dense exospore. the polar sac is cup-like and lacks a layered anchoring disc. the polar filament is arranged in two to three isofilar coils in the half of the spore opposite the nucleus. the coupling between the polar sac and the polar filament is characteristic. the surface of the polar filament is covered with regularly arranged membraneous chambers resembling a honeycomb. There is no polaroplast of traditional type. the cytoplasm lacks polyribosomes. the nucleus has a prominent, wide nucleolus. the two spore types have identical construction, but differ in dimensions and electron density. Free living spores are about 3.2 μm wide, the diameter of the polar filament proper is 102-187 nm, the chambers of the honeycomb are 70-85 nm high, and the polar sac is up to 425 nm wide. Living spores in the vesicle-bound sporogony are about 2.1 μm wide, the polar filament measures 69-102 nm, the chambers of the honeycomb are about 45 nm high, and these spores are more electron dense. Comparisons of cytology (especially the construction of the spore wall and the polar filament and associated structures) and life cycles reveal prominent differences among the Chytridiopsis-like microsporidia, and close relationships between the families Chytridiopsidae and Metchnikovellidae.     

枣树体细胞胚发生和组织学研究

以临泽小枣子叶切块为外植体,在附加０．２mg/L IBA 1．0mg/L 6-BA的ＭＳ培养基上１周后切块边缘可诱导出白色胚愈伤组织,继续培养１个月后愈伤组织中产生体细胞胚。体细胞胚发生不同步,经历球形胚、心形胚、子叶胚等阶段,与合子胚发育途径相似。组织切片表面胚性愈伤组织细胞体积小,细胞核大、细胞质浓,细胞排列紧密;而非胚性愈伤组织细胞体积大、细胞核小、细胞质稀薄,子叶胚时期体细胞胚内部出现维管束,并观察到螺纹导管。     

The ultrastructure of the sensory cells in the chemoreceptor of the ommatophore of Helix pomatia L.

Most of the sensory cells found in the chemoreceptor of the ommatophore of Helix pomatia are typical bipolar cells. The chemoreceptor is deveded by a furrow into two parts; within the ventral subdivision the layer of sensory cell bodiesis thicker than in the dorsal part. According to the differentiations of the apical surface of the dendrites, it is possible to distinguish six different classes: a) dendrites with one cilium and 75 nm thick cytofila (sometimes dendrites of identical appearance posses more than one cilium); b)dendrites with several cilial and 150 nm thick cytofila; c) dendrites with several cilia, 50 nm thick cytofila, and long, striated rootlets; d) dendrites with several cilia bur without cytofila; e) dendrites with 130 nm thick cytofila but without cilia; and f) dendrites with 65 nm thick cytofila but without cilia; dendrites of this class are the only ones with a cytoplasm more electron dense than that of the surrounding supporting cells. All these dendrites are connected to the surrounding supporting cells by terminal bars, each consisting of zonula adhaerens, aonula intermedia and zonula septata. The perikarya of the sensory cells measure approximately 15 mum by 8 mum and enclose 10 mum by 6 mum large nuclei. Axons, originating from these perikarya, extend to the branches of the digital ganglion. In the distal part of this gangloin the axons come into synaptic contact with interneurons, but in our electron micrography it was not possible to coordinate processes and synapses with the corresponding neurons.     

Ultrastructure of muscle cells in the adductor of the boring clam Tridacna crocea

The ultrastructure of the adductor muscle of the boring clam (Tridacna crocea) was investigated. The adductor was composed of opaque and translucent portions. The opaque portion contained smooth muscle cells; the translucent portion contained obliquely striated cells. Smooth muscle cells were classified, according to the statistically analyzed diameters of their thick myofilaments, into two types, S-1 and S-2. S-1 cells had thick myofilaments, 50–60 nm in diameter. S-2 cells had thick myofilaments of two sizes, about 55–65 nm and 85–100 nm in diameter, respectively. Obliquely striated muscle cells in the translucent portion were also classified into two types: O-1 cells, with thick myofilaments 30–35 nm in diameter, and O-2 cells, with myofilaments of 50–60 nm.     

Rotary shadowing of collagen monomers, oligomers, and fibrils during tendon fibrillogenesis

Collagen monomers, oligomers, and fibrillar structures were isolated from chick tendons at various stages of development and studied by rotary shadowing. Monomers of Type I collagen, solubilized in 0.15 M NaCl solutions, were mostly present as collagen, pN-collagen, and pC-collagen with few procollagen molecules. They did not form polymers, nor were they associated with a carrier. Dimers of fibrillar collagen molecules were arranged in a 4-D stagger, suggesting that this was the preferred molecular interaction for the initiation of collagen fibrillogenesis. Type XII collagen molecules were mostly free, but some were attached by their central globular domain to one end of free fibrillar collagen molecules. Tenascin and Type VI collagen were also identified. The fibril populations consisted of collagen and beaded structures. These fibrils consisted of beads (globular domains) about 23 nm in diameter, separated by a period about 27 nm in length. Beads were linked by filamentous structures. These beaded fibrils probably represent the microfibrils of elastin.