Liver ultrastructure in sheep fasciolosis

This experiment has studied the changes in the liver in sheep during experimental infection with fascioliasis. Sheep were infested with 300 adoliskaria and observed changes up to 142 days. At the ultrastructural level in animals with fascioliasis, destructive changes were found in the liver. In the nucleus, the number of nucleoli decreases or they disappear completely, fragmentation of heterochromatin occurs and the content of euchromatin increases. The karyolemma exfoliates from the nucleus, as a result of which the perinuclear space expands. Mitochondria swell, they are polymorphic, and have an electron-dense matrix. At the beginning of the invasion, their number increases, and then their vacuolization, destruction occurs, while under the action of anthelmintic drugs and fasciolus toxins, the structure of the endolasmic network first of all changes: its cavities expand, then fragmentation occurs. Ribosomes are sprayed into the cytoplasm of the hepatocyte. It should be noted that under the influence of hexichol, acemidophene and thiopagol in the liver, membrane structures change most significantly and undergo necrobiosis.

Ultrastructural lesions in the myocardium and kidneys of rabbits in experimental acute Amorimia exotropica poisoning

Amorimia exotropica is an important plant associated with sudden death in cattle in Southern Brazil. In order to understand the mechanisms by which A. exotropica causes acute lesions in the heart and kidney of intoxicated animals, an experiment was conducted to determine the histopathology and ultrastructure of myocardial and renal lesions of intoxicated rabbits. After receiving 18g/kg of dried plant, six rabbits died suddenly. At necropsy, the liver was swollen and no other macroscopic lesions were observed. Histologically, centrolobular and midzonal hepatocytes were vacuolated. These vacuoles were strong PAS stained positive, suggesting that they corresponded to glycogen accumulations. In some regions of the ventricular septum and ventricles were found vacuoles of different sizes and the kidneys of two rabbits showed vacuolar degeneration on distal convoluted tubules. Ultrastructurally, the myocardium had cardiomyocytes swelling with separation of myofibrils bundles and rupture and disorganization of the sarcomeres. The mitochondria displayed swelling, disorganization, disruption of the mitochondrial cristae, and electron-dense matrix. Some mitochondria exhibited eccentric projections of their membranes with disruption of both outer and inner membranes. The sarcoplasmic reticulum had no alterations, whereas the T-tubule system was occasionally dilated and ruptured. The kidneys had mitochondrial swelling with disorganization and disruption of the mitochondrial cristae. The vacuoles result from the swelling of the endoplasmatic reticulum and usually were located between two basolateral infoldings and mitochondria, occurring preferentially around the nucleus. The myocytes and T system damages induced by A. exotropica result in acute heart failure and death. Furthermore, this mechanism of cardiotoxicity may be common to all plant containing monofluoroacetate.

Fine structure of the anterior adhesive apparatus (head organs) of Bravohollisia gussevi Lim, 1995 (Monogenea: Ancyrocephalidae)

A study of the anterior adhesive apparatus (head organs) of Bravohollisia gussevi Lim, 1995 was carried out using light and electron microscopy. The anterior adhesive apparatus or head organs in B. gussevi comprise 6 circular openings or apertures in the antero-lateral region, associated pits lined with specialized microvillous tegument that differ from the general body tegument, a bundle of ducts, and uninucleate gland cells located lateral to the pharynx. The uninucleate glands of the anterior adhesive apparatus (head organs) comprise 2 types of cells, one kind of cell producing rod-like bodies (S1) and the other oval bodies (S2). The S1 bodies are filled with numerous, less electron-dense vesicles in an electron-dense matrix, while S2 bodies have no vesicles but contain a more homogeneous electron-dense matrix. Interlinking band-like structures were observed between S1 bodies. Similar band-like structures were found between S2 bodies. The formation of S1 bodies was followed by transmission electron microscopy. However, the formation of S2 bodies was unclear and could not be resolved. Uniciliated structures were also observed around the openings of the anterior adhesive apparatus. Each uniciliated structure is usually associated with an opening of a gland cell producing granular, electron-dense, secretory bodies, which differ from the secretions produced by the lateral gland cells of the anterior adhesive apparatus.

Sporidial mating and infection process of the smut fungus, Ustilago hordei, in susceptible barley

Ustilago hordei (Pers.) Lagerh. causes covered smut of barley and oats. Sporidial mating and the infection process on compatible barley plants, cv. Hannchen, were investigated using light microscopy and scanning and transmission electron microscopy. Within 2 h after mixing of sporidia of opposite mating types on water agar, polar conjugation tubes emerged that subsequently fused, producing infection hyphae at the junctions. Similar events occurred on germinated barley shoots, although sporidia regularly produced several conjugation tubes, of which only one was involved in mating. Tubes emerging from the sides of cells were also observed. Infection hyphae emerged from either the conjugation tube or conjugated cell body. Hyphae elongated along the shoot surface until characteristic crook and appressorium-like structures were formed. An invading hypha emerged beneath this structure and directly penetrated the underlying epidermal cell. Hyphae extended both intra- and inter-cellularly into tissues, without much branching, before becoming established in the shoot meristematic region. Plant plasma membranes remained intact during pathogen ingress and an electron-dense matrix of unknown origin appeared in the interface between plant plasma membrane and invading hypha. A large fungal biomass was generated in the host spike tissue at 42–63 days postinoculation during the development of the floral meristem.Key words: Hordeum vulgare, pathogen, sporidia, teliospores, ultrastructure, Ustilaginales.

An ultrastructural study of pollen development in tomato (Lycopersicon esculentum). I. Tetrad to early binucleate microspore stage

Microspores undergo considerable ultrastructural changes between the tetrad and early binucleate microspore stages of microsporogenesis in tomato (Lycopersicon esculentum). Pollen wall deposition began late in the tetrad stage, and by the early microspore stage a lamellar foot layer and tectum were deposited. Sculpturing of the tectum was evident by the early binucleate microspore stage. Dictyosomes and vesicles were abundant during the period of pollen wall formation. Plastids were associated with the endoplasmic reticulum (ER) to form plastid–ER complexes, from the late tetrad to the vacuolate microspore stage. At the vacuolate microspore stage, endoplasmic reticulum independent of plastids was also observed, and at the early binucleate microspore stage ER was not associated with plastids. Free ribosomes were evenly distributed throughout the cytoplasm until the vacuolate microspore stage when they were organized into polysomes. Mitochondria were spherical to ellipsoid, with an electron-dense matrix and swollen cristae, until the early binucleate microspore stage when they were highly elongate and became convoluted. Key words: Lycopersicon esculentum, microsporogenesis, pollen development, tetrads, tomato, ultrastructure.

Development and Ultrastructure of Primary Secretory Ducts in the Stem of Semecarpus Anacardium (Anacardiaceae)

Development and ultrastructure of primary phloem secretory ducts in the stem of Semecarpus anacardium are described. Ducts are formed schizolysigenously in the phloic procambium of the young stem just below the shoot apex. The cytoplasm of the epithelial cells is rich in osmiophilic material (resin), plastids, rough endoplasmic reticulum (RER), free ribosomes, polysomes, mitochondria with swollen cristae and golgi bodies. The plastids are of varying shapes, have an electron dense matrix and a poorly developed internal membrane system. They are highly contorted and invariably surrounded by a sheath of RER. Osmiophilic material is frequently observed in the plastid stroma. Epithelial cell cytoplasm and its plastids are the plausible sites of resin formation. The gum component of the secretion may be derived from the inner tangential wall (ITW). The secretory substances reach the apoplast principally by granulocrine and eccrine secretion. Epithelial cells are linked with each other and with the neighbouring cells by numerous plasmodesmatal connections. Some epithelial cells undergo lysis while others retain their secretory function.

Evolutionary conservation of a microbody targeting signal that targets proteins to peroxisomes, glyoxysomes, and glycosomes.

Peroxisomes, glyoxysomes, glycosomes, and hydrogenosomes have each been classified as microbodies, i.e., subcellular organelles with an electron-dense matrix that is bound by a single membrane. We investigated whether these organelles might share a common evolutionary origin by asking if targeting signals used for translocation of proteins into these microbodies are related. A peroxisomal targeting signal (PTS) consisting of the COOH-terminal tripeptide serine-lysine-leucine-COOH has been identified in a number of peroxisomal proteins (Gould, S.J., G.-A. Keller, N. Hosken, J. Wilkinson, and S. Subramani. 1989. J. Cell Biol. 108:1657-1664). Antibodies raised to a peptide ending in this sequence (SKL-COOH) recognize a number of peroxisomal proteins. Immunocryoelectron microscopy experiments using this anti-SKL antibody revealed the presence of proteins containing the PTS within glyoxysomes of cells from Pichia pastoris, germinating castor bean seeds, and Neurospora crassa, as well as within the glycosomes of Trypanosoma brucei. Western blot analysis of purified organelle fractions revealed the presence of many proteins containing this PTS in both glyoxysomes and glycosomes. These results indicate that at least one of the signals, and therefore the mechanism, for protein translocation into peroxisomes, glyoxysomes, and glycosomes has been conserved, lending support to a common evolutionary origin for these microbodies. Hydrogenosomes, the fourth type of microbody, did not contain proteins that cross-reacted with the anti-PTS antibody, suggesting that this organelle is unrelated to microbodies.

THE SECRETORY PATHWAY IN PLATELETS STUDIED BY CRYO-FIXATION

It is widely held, that the constituents packed in the a -granules are released by stimulated platelets via the surface connected system (SCS). By means of the fast-freezing and freeze substitution technique (which allow the investigation of membrane fusion) we found a secretory pathway in platelets (compound exocytosis) without an involvement of the SCS during the release of a-granules. To study the process of a-granule secretion human platelets concentrated in citrated blood plasm were stimulated with thrombin or collagen. 20 - 120 seconds after stimulation the platelets were rapidly frozen with a metal-mirror attachment to the KF 80 cryofixation unit (REICHERT-JUNG). Using plastic spacers droplets of the PRP were slammed against a copper block at 80 K at a rate of 0.2 m/sec. After cryofixation the specimens were transferred (in liquid nitrogen) into a Cs-auto cryosubstitution unit (REICHERT-JUNG). Cryosubstitution was programmed for 48h at 193 K in acetone with 4% osmium tetroxide. The temperature went automatically up to room temperature at a rate of 10 K/h. The specimens were embedded in araldite. The analysis of serial ultrathin sections of platelets in different phases of exocytosis revealed the following. a -granules in apposition showed different stages of swelling and dispersal of their electron dense matrix. Membrane appositions were also found between a -granules. The contraction of a sphere of microfilaments and microtubules during stimulation seemed to support this process. On the other hand this internal contraction prevented most of the a-granules from contacting with the plasmalemma. We observed fusion between swollen -granules in apposition and the plasmalemma and swollen and unswollen a -granules. Thus, large compound granules were formed frequently before fusion of the secretory organelles with the plasmalemma took place. These observations suggested that a -granules in stimulated platelets performed a compound exocytosis after swelling. The process seemed to start with the apposition of a -granule membranes to the plasmalemma. It cannot yet be answered whether the swelling of the granules is due to an osmotically driven influx of water or due to an influx after microfusion.Supported by DFG, Grant Mo 124/2-4

Cytochemical and enzymatic characterization of the sporulation septum of Streptomyces antibioticus

The sporulation septum in Streptomyces antibioticus is formed by two thin electron-dense layers which constitute the annuli, separated by a thick and moderately electron-dense matrix which forms the deposits. By means of cytochemical techniques and enzymatic digestion with lysozyme, we have shown that deposits and annuli have a different chemical composition. Deposits are formed by peptidoglycan and are highly sensitive to lysozyme, whereas annuli, probably due to localized chemical modifications of the peptidoglycan or high degrees of peptide cross-linking, are lysozyme resistant. Our results suggest that the deposits, unlike the annuli, are only temporary assisting structures which are degraded during spore maturation.