A LIGHT AND ELECTRON MICROSCOPE STUDY OF INTERLAMELLAR POLYGLUCOSIDE BODIES IN OSCILLATORIA CHALYBIA

A light and electron microscopic study of interlamellar granules in Oscillatoria chalybia was made to determine their physiological nature. Oscillatoria chalybia was cultured under continuous light in media of high nitrogen content, moderate nitrogen content and low nitrogen content. Cultures growing vigorously in a medium of moderate nitrogen content were placed in darkness for an additional 96 hr. Periodic acid-Schiffs reagent tests were made on specimens from these 4 cultural conditions. Electron microscopic studies of interlamellar granules were correlated with the cytochemical tests. It is shown that diastase digestion will eliminate the PAS-positive substance and the interlamellar granules. Conclusions are that the interlamellar granules are polyglucoside in nature and that they vary in number and size with available nitrogen, light intensity and age of culture in such a way as to indicate that they are food reserves.     

The effect of food odor background on gustatory preferences and gustatory behavior of carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> and cod <Emphasis Type="Italic">Gadus morhua</Emphasis>

It was shown that stimulation by food odor (aquatic extract of food organisms, 10^?2 and 10^?3 g/l) does not cause shifts in gustatory preferences in carp Cyprinus carpio and cod Gadus morhua but modifies gustatory behavior. The level of consumption by carp of control granules and granules with attractive, by taste, L-proline (0.1 M) or deterrent L-lysine (0.1 M) (item by item presentation of granules) and by cod of control granules and granules with indifferent, to it, L-asparagine (0.1 M) (presentation of 10 granules simultaneously) is similar prior to and during olfactory stimulation. In the presence of food odor, the duration of taste testing for most types of granules, as well as the number of repeated graspings of granules with an attractive taste do not change in fish. At the same time, granules with indifferent or repulsive gustatory properties are rejected and repeatedly grasped by fish against the background of food odor more frequently than in water without odor. Olfactory stimulation leads to a considerable increase in the average number of graspings per one grasped granule with an indifferent or repulsive taste. Such behavior manifested by fish in the presence of food odor in response to granules with unattractive gustatory properties is apparently caused by the contradiction between the information coming via different chemosensory canals—olfactory and gustatory. The obtained results indicate that food stimulation caused by food odor in nature can lead to an increase in the actual consumption of only those accessible food items that have an attractive taste for fish.     

Pesta granule trials with Aspergillus alliaceus for the biocontrol of Orobanche spp.

The pathogenic fungus Aspergillus alliaceus has been shown to have potential for the biocontrol of Orobanche spp. (broomrape), a root parasitic plant. The effectiveness of A. alliaceus in reducing Orobanche infection was analysed using pesta granules prepared with different food formulations. The results showed that pesta granules comprising of fungal mycelia/spore mixtures from liquid and solid culture, sclerotia and fungal mycelia reduced Orobanche infection to a greater extent in below ground conditions when applied early and at high doses before crop sowing. In addition, pesta granules eliminated the risk of broomrape contamination within a 0.2–0.3 cm diameter of the granules. The sclerotial pathogenicity of A. alliaceus was compared with those of other fungi reported in other studies. In addition, some morphological and histological studies on the fungal pathogenicity on broomrape plants after infection are presented. The present study reveals the potential of sclerotial A. alliaceus pesta granule applications for long-term broomrape biocontrol under field conditions.     

The distribution of zinc, cadmium, lead and copper within the hepatopancreas of a woodlouse

The distribution of metals within the hepatopancreas of Oniscus asellus (Crustacea, Isopoda) from two uncontaminated sites, and two sites contaminated with zinc, cadmium and lead, has been studied by atomic absorption spectrophotometry, light microscopy, transmission and scanning electron microscopy and X-ray microanalysis. The hepatopancreas contains two types of intracellular granule. The first type, in the S cells, are spherical granules which contain copper, sulphur and calcium. In woodlice from contaminated sites, these ‘copper’ granules, also contain zinc, cadmium and lead. The second type, in the B cells, are flocculent deposits which contain iron. In woodlice from contaminated sites, these ‘iron’ granules also contain zinc and lead. Moribund woodlice from contaminated sites have large numbers of ‘copper’ and ‘iron’ granules in the hepatopancreas and a fine deposit of zinc and lead on the membranes of the cells. There are numerous microorganisms in close association with the microvillous border of the hepatopancreas of woodlice from all four sites. Within the microorganisms of Oniscus asellus from contaminated sites, there are deposits of material which contain zinc, lead, calcium and phosphorus ‘Copper’ and ‘iron’ granules could have evolved as storage sites for essential metals to be utilized when demand from the body exceeds uptake from the food. Woodlice in contaminated sites may be able to ‘detoxify’ potentially harmful amounts of essential and non-essential metals by storing them in a relatively insoluble form within these granules.     

Histophysiology of digestion and observations on the structure of the alimentary canal in the ectosymbiont Chaetogaster limnaei limnaei Baer, 1827 (Annelida: Oligochaeta)

The naidid oligochaete Chaetogaster limnaei limnaei has an alimentary canal consisting of a mouth, pharynx with a dorsal pharyngeal pad, esophagus, stomach, anterior and posterior intestine, and anus. The diet is omnivorous but limited by particle size. Unattached food organisms are sucked into the pharynx while sessile organisms are plucked from the substratum. Granules of acid mucosubstances that stain purple with neutral red are secreted into the stomach lumen after food enters, rapidly increasing the acidity from pH 3 to 1.5. Acid induced lysis of the organisms initiates autolysis before the food is passed into the alkaline, pH 7 to 8, anterior intestine. Ciliated intestinal cells showed arylamidase, acid phosphatase and C-esterase active granules indicating primary lysosomes with secondary lysosomes being recognized in electron micrographs suggesting intracellular digestion. Arylamidase and alkaline phosphatase activity appears in the intestinal margins during the alkaline phase of digestion. Scattered, pyramidal cells found only in the anterior intestine contain yellow refractile spheres. The spheres stain alcian blue pH 2.5 and bromophenol blue positive and exhibit a strong acid phosphatase activity all the time with A-esterase active granules surrounding them. Glycogen and lipids are stored mainly in the chlorogague cells. Many of the yellow refractile granules in the stomach and intestinal cells are bacteria.     

Ultrastructural and Fluorescence Microscopical Characterization of the Intestinal Endocrine Cells in a Cyclostome,Myxine glutinosa

Endocrine cells of so-called basal-granulated-open type in the intestinal epithelium of a cyclostome, the Atlantic hagfish (Myxine glutinosa), are characterized ultrastructurally and fluorescence microscopically. These cells regularly extend from the basal lamina to the gut lumen, ending in an apical process with microvilli and a filamentous surface coat. Fasting results in an accumulation of secretion granules in all cytoplasmic portions, except for the terminal web area. A similarity is recorded between the distribution of secretion granules and the finely granular fluorescamine-induced fluorescence, suggesting that the fluorescence is associated to some component(s) of the secretory granules. Granule release may take place at the base after an adequate stimulus (presence of food?) at the luminal portion of the cells. The formaldehyde condensation technique shows that insulin-containing hagfish islet parenchymal cells, but not intestinal endocrine cells, store dopamine after intestinal supply of the amine precursor. Acidification of formaldehyde vapour-fixed intestinal epithelium induces fluorescence in the granules of zymogen cells but not of endocrine cells, indicating a low concentration of tryptophyl-peptide(s) in the secretory granules of hagfish intestinal endocrine cells.     

Transmission and scanning electron microscopic study of adepidermal granules of teleosts and amphibia

Summary Granules in the adepidermal space of larvae of Salmo irideus, Hynobius tokyoensis and Rhacophorus buergeri, were observed by transmission and scanning electron microscopy. Adepidermal granules of S. irideus were smooth and spherical structures, those of H. tokyoensis were smooth and spherical, or oval, while in R. buergeri these granules appeared as single or grouped tangled strand-like or starfish-like structures under the scanning electron microscope. These adepidermal granules were spread all over the basal lamina in every animal investigated. The different sizes of adepidermal granules of S. irideus and H. tokyoensis seen under the transmission electron microscope are not the result of differently sectioned faces of granules, but the granules themselves exhibit different sizes. The probable functions of these granules are discussed.     

Ultrastructural cytochemistry of glycoconjugates in basophils from humans and animals

Summary The distribution of glycoconjugates in the basophil granules of humans, guinea pigs, and rabbits was compared. The observation of acid mucopolysaccharides using the dialyzed iron method and of sulfated glycoconjugates using the high iron diamine method revealed three types of reactions in the basophil granules of all three species: (1) granules showing a strong overall reaction, (2) granules showing reaction only at their periphery, and (3) granules showing no reaction. With regard to the relationship between maturation and the types of basophil granules, it appeared that, in general, there were many type-1 granules among immature basophils, but that these granules decreased in mature basophils as type-3 granules increased. The reaction patterns of periodate-reactive neutral glycoconjugates, as shown by the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method, were different from those of acid mucopolysaccharides: the reaction of basophil granules was diffusely positive, and localization at the periphery was rarely observed. Therefore, unlike the acid mucopolysaccharides, it was difficult to classify the glycoconjugates into three types. However, as with acid mucopolysaccharides, there was a tendency for periodate-reactive glycoconjugates to decrease as maturation progressed. In terms of different species of animals, the reaction of periodate-reactive glycoconjugates with PA-TCH-SP was stronger in humans and rabbits than in guinea pigs.     

The ultrastructure of guinea pig heterophil granulocytes and the heterogeneity of the granules

Summary The development of the heterophil granulocytes in the bone marrow of the guinea pig is described. During the maturation of these cells, three types of granule are formed, not only the azurophil and specific granules already described in other mammals but also a third type of granule referred to here as the nucleated granule. During the process of maturation of the cells, these three types of granule are formed successively. On this basis, two steps can be distinguished in the promyelocyte phase in which primary (nucleated and azurophil) granules are formed, i.e. an early and a late stage, nucleated granules being formed in early and azurophil granules in late promyelocytes. Secondary (specific) granules occur first in myelocytes. In mature heterophils of the guinea pig the granule population is composed of about 85% secondary granules, about 10% azurophil granules, and about 5% nucleated granules. The changes in the granule population during the maturation process were quantified. The observations and calculations point to the occurrence of three mitoses: one in the early and one in the late promyelocyte and the third in the myelocyte.     

Co-localization of the adipokinetic hormones I and II in the same glandular cells and in the same secretory granules of corpus cardiacum of Locusta migratoria and Schistocerca gregaria

Summary The immunocytochemical reactivity of the glandular cells of the corpus cardiacum (CCG-cells) of Locusta migratoria and Schistocerca gregaria was investigated at the electron-microscopic level, using the protein A-gold method, with three antisera against fragments of the adipokinetic hormones AKH I and AKH II. This combination of antisera permitted discrimination between anti-AKH I and anti-AKH II immunoreactivity. Fixation in a mixture of 2% glutaraldehyde and 2% formaldehyde, in combination with low-temperature embedding in Lowicryl K₄M, produced the highest and most consistent selective immunogold labelling of the secretory and ergastoplasmic granules. All secretory granules in all CCG-cells investigated possessed a distinct anti-AKH I-immunopositive reaction, whereas most secretory granules showed a weaker anti-AKH II immunoreaction. Ergastoplasmic granules reacted similar to the secretory granules. The average immunolabelling of the secretory granules was higher in the processes than in the cell bodies of the CCG-cells. The results in Schistocerca gregaria were essentially similar to those in Locusta migratoria. It is concluded that (i) the individual CCG-cells synthesize AKH I as well as AKH II; (ii) these hormones coexist in the same ergastoplasmic and secretory granules; and (iii) these granules contain a higher content of AKH I than AKH II.     

Graniferous tracheary elements in the haustorium ofAtkinsonia ligustrina,a root hemi-parasite of theLoranthaceae

Summary The xylem conducting cells that form the vascular core in the root haustorium ofAtkinsonia ligustrina are shown to be graniferous tracheary elements. Their luminal contents occur in the form of granules, amorphous bodies or finely dispersed material derived from the granules. Cells of this type are known for theSantalaceae, but have not previously been reported unequivocally in theLoranthaceae. The matrix of the granules consists of compacted fibrillar material often with the central region of the granule showing a crystalline substructure. The granules stain positively for protein. In some instances the matrix may become secondarily impregnated with phenolic compounds. The granules arise during differentiation of the tracheary element and develop as accumulations of matrix within dilated cisternae of endoplasmic reticulum. The function of graniferous tracheary elements inAtkinsonia may be to help regulate the flow of xylem sap from host to parasite, as has been suggested for theSantalaceae.     

Characterization of a Potato Starch-digesting Bacterium and Its Production of Amylase

A bacterium which can utilize potato starch granules as sole carbon source was isolated and identified as Bacillus circulans from its physiological and biochemical properties. Scanning electron microscopic observation of potato starch granules recovered from the culture broth revealed that granules were degraded gradually from their surface resulting in elongated granules with layered structures on their surface. This bacterium produced extracellular amylase which can digest potato starch granules in vitro. The amylase has a unique property in that it produces only maltohexaose from gelatinized starch in the early stage of the reaction. For the production of this amylase potato starch was found to be most effective while soluble sugars including gelatinized starch and maltose had little effect.     

Heterogeneity of the blood group ABH antigens and variation in the expression of these antigens of secretory granules in human cervical glands

Summary Cytochemical localization of blood group ABH antigens was examined in secretory cells of human cervical glands by application of a post-embedding lectin-gold as well as immuno-gold labeling procedure using monoclonal antibodies. Blood group specific lectins such as Dolichos biflorus agglutinin (DBA), Helix pomatia agglutinin (HPA), Griffonia simplicifolia agglutinin I-B₄ (GSAI-B₄) and Ulex europaeus agglutinin-I (UEA-I) reacted with secretory granules but not with other cytoplasmic organellae such as nucleus and cell membrane. The reactivity of secretory granules with these lectins showed strict dependence on the blood group and secretor status of tissue donors. The binding patterns with these lectins were not homogeneous, but exhibited marked cellular and subcellular heterogeneity. Thus, for example, in blood group A individuals, some granules were stained strongly with DBA and others were weakly or not at all with the lectin. Such a heterogenous labeling with the lectin was observed even in the same cells. Similar results were obtained with UEA-I and GSAI-B₄ staining in blood group O and B secretor individuals, respectively. Monoclonal antibodies likewise reacted specifically with the granules but they occasionally bound to some nucleus. The labeling pattern of the antibodies with the granules was essentially the same as those of lectins. However, difference was also observed between monoclonal antibody and lectin staining, that is, monoclonal anti-A antibody reacted weakly but consistently with granules from blood group A nonsecretors but DBA (HPA) did not; staining with UEA-I was observed in granules from the secretor individuals of any blood groups whereas monoclonal anti-H antibody reacted with granules from blood group O and some A secretor individuals but not from B and AB secretor individuals; GSAI-B₄ reacted uniformly with granules throughout the cells whereas monoclonal anti-B antibody bound to limited number of granules in the same cells. This was confirmed by the double labeling experiments with the lectin and the antibody. These results suggest that the different types of antigens as to the binding ability for monoclonal antibodies and lectins are expressed on different granules in the same cell.     

A Comparative Study of the Storage Carbohydrate Granules from Euglena (Euglenida) and Pavlova (Prymnesiida)1,2

ABSTRACT The storage carbohydrate granules from Euglena and Pavlova were compared by light and electron microscopy. Freezeetch studies demonstrated that while both types of granules are crystalline, they have different structures. The elemental microfibril of the euglenoid granule measures 4 nm, and the elemental striation of the granule from Pavlova is 22 nm. The granules each have a unique X-ray diffraction pattern. The storage carbohydrate granules from Pavlova are not the same as paramyton, and the term “paramylon” should be reserved for the euglenoid storage carbohydrate.     

Amebic Parasites of Fishes*

SYNOPSIS. Entamoeba molae n. sp. was found in the hindgut of Mola mola collected from the coast of southern California. The trophic stages are actively motile and contain numerous food vacuoles. The nuclear peripheral chromatin is usually conspicuous. A subperipheral crescent of granules is the most distinctive feature of the nucleus. The endosome is eccentric and often surrounded by a periendosomal ring. Measurements: cell body 16.4 × 22.9 μ nucleus 4.8 μ in diameter. Cysts were not seen. We believe that few amebas have been described from fishes because few fishes have been examined for amebas.     

Expression of an amylosucrase gene in potato results in larger starch granules with novel properties

Main conclusion

Expression of amylosucrase in potato resulted in larger starch granules with rough surfaces and novel physico-chemical properties, including improved freeze–thaw stability, higher end viscosity, and better enzymatic digestibility. Starch is a very important carbohydrate in many food and non-food applications. In planta modification of starch by genetic engineering has significant economic and environmental benefits as it makes the chemical or physical post-harvest modification obsolete. An amylosucrase from Neisseria polysaccharea fused to a starch-binding domain (SBD) was introduced in two potato genetic backgrounds to synthesize starch granules with altered composition, and thereby to broaden starch applications. Expression of SBD–amylosucrase fusion protein in the amylose-containing potato resulted in starch granules with a rough surface, a twofold increase in median granule size, and altered physico-chemical properties including improved freeze–thaw stability, higher end viscosity, and better enzymatic digestibility. These effects are possibly a result of the physical interaction between amylosucrase and starch granules. The modified larger starches not only have great benefit to the potato starch industry by reducing losses during starch isolation, but also have an advantage in many food applications such as frozen food due to its extremely high freeze–thaw stability.     

Starter culture of <Emphasis Type="Italic">Pseudomonas veronii</Emphasis> strain B for aerobic granulation

Aggregation of bacterial cells is used in formation of microbial granules. Aerobically grown microbial granules can be used as the bio-agents in the treatment of wastewater. However, there are problems with start up of microbial granulation and biosafety of this process. Aim of this research was selection and testing of safe microbial strain with high cell aggregation ability to shorten period of microbial granules formation. Five bacterial strains with cell aggregation index higher than 50% have been isolated from the granules. Strain of Pseudomonas veronii species was considered as most probably safe starter culture for granulation because other strains belonged to the species known as human pathogens. The microbial granules were formed after 3 days of cultivation in case when P. veronii strain B was applied to start-up aerobic granulation process using model wastewater. The granules were produced from activated sludge after 9 days of cultivation. Microbial aggregates produced from starter culture of P. veronii strain B were more compact (sludge volume index was 70 ml/g) than those produced from activated sludge (sludge volume index was 106 ml/g). It is a first proof that application of selected safe starter pure culture with high cell aggregation ability can accelerate and enhance formation of microbial granules.     

Gelatinase Granules of Neutrophil Granulocytes

The neutrophil contains numerous granules of various composition and structure. For decades, the neutrophil was believed to contain only two granule types, peroxisomes, or peroxidase-positive granules, and peroxidase-negative granules. Later, existence of the third type distinguished by the presence of gelatinase hydrolyzing collagen and gelatin was proposed. Gelatinase was found in the granules that are lighter as compared to the common peroxidase-negative granules and represent their subpopulation. In addition to gelatinase, these granules contain beta-2 microglobulin, cytochrome b ₅₅₈, as well as receptor and adhesion proteins. Upon stimulation by inflammatory mediators, the gelatinase granules are secreted before the common peroxidase-negative granules. Their exocytosis mediates delivery of new adhesion proteins to the plasma membrane, which is required for maintenance of permanent and fast cell adhesion to the endothelium. The released gelatinase allows the neutrophil to penetrate through the basement membrane of the endothelium.     

A new type of ribonucleoprotein constituent of the polytene nucleus of the salivary glands ofChironomus thummi andCh. tentans

Using electron spectroscopic imaging, a new type of small granular structural constituent has been observed in the extrachromosomal zone of the polytene nucleus of the salivary gland cells ofChironomus thummi andChironomus tentans. These granules appear isolated or in small clumps and are often seen to be connected with surrounding thin fibrils. They are stained by the EDTA procedure, which is preferential for nuclear ribonucleoprotein (RNP) constituents, and by the bismuth oxynitrate method for visualizing phosphorylated compounds. The granules are 15–23 nm in diameter and are digested by prolonged post-embedding RNAse hydrolysis. These structural elements contain the highest concentration of phosphorus in the interchromosomal space as revealed by electron energy loss spectroscopy. The small granules exhibit several morphological and cytochemical features in common with interchromatin granules, but they are not labeled with antibodies directed against extranucleolar small nuclear RNPs (snRNPs), as are interchromatin granules.