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S F Conti

Publications and source records attributed to S F Conti.

At least 37 records · Page 2Linked to original sources

Characterization of germination and activation of Bdellovibrio bdellocysts.

A simple method of assaying germination of bdellocysts in liquid medium has been devised. Bdellocysts can be induced to germinate by any of eight L-amino acids or the monovalent cations K+ and NH4+. L-Glutamine was the best individual inducer of germination, although the resulting rate of germination was much slower than in a complex medium. The use of a defined germination medium containing L-glutamine, KCl, and NH4Cl produced a faster rate of germination than did complex media. Bdellocysts germinated most rapidly at pH 8.0 and at 35 degrees C and required aerobic conditions. Respiration of bdellocysts began to increase at 3 min after the addition of germinants. Germination was inhibited by respiratory-chain inhibitors and by inhibitors of macromolecular synthesis. When bdellocysts were heat shocked at sublethal temperatures for short periods, there was no effect on the rate of germination in the defined germination medium or in the complex medium. However, heat-shocked bdellocysts germinated at a much faster rate in the presence of single inducers of germination when compared to nonshocked bdellocytes.

Amino Acids↗

Chemotaxis toward amino acids by Bdellovibrio bacteriovorus.

Chemotaxis toward amino acids by Bdellovibrio bacteriovorous strain UKi2 was studied by the capillary technique of Adler (J. Gen. Microbiol. 74:77-91, 1973). Chemotaxis was shown to be optimal when the capillaries were incubated at between 15 and 40 degrees C for 30 min; the optimal pH was between 7.0 and 8.2. The chemotactic response was proportional to the density of the suspension of bdellovibrios up to a density of 10(8) cells/ml. B. bacteriovorus was attracted to L-asparagine, L-cysteine, L-glutamine, glycine, L-histidine, L-lysine, and L-threonine. The possible roles of chemotaxis in the life of B. bacteriovorus are discussed.

Amino Acids↗

Characterization of bdellocysts of Bdellovibrio sp.

Bdellovibrio sp. strain W will infect and produce resting cells, termed bdellocysts, in a variety of gram-negative bacteria. Bdellocysts appeared to be produced only within susceptible prey and never in their absence. Optimum conditions for encystment included infection of stationary-phase prey cells in 0.05 M potassium phosphate buffer (pH 7.5) at concentrations of prey and bdellovibrios of 2 X 10(9) cells per ml with a multiplicity of infection of unity. Bdellocysts contained more deoxyribonucleic acid, ribonucleic acid, protein, and carbohydrate per cell than did vegetative cells. Poly-beta-hydroxybutyrate and dipicolinic acid were not detected. Bdellocysts were more resistant than vegetative cells to effects of elevated temperatures, sonic treatment, and desiccation. Bdellocysts remained viable for extended periods when incubated in the absence of prey, whereas vegetative cells lost viability rapidly under the same conditions. Their survival under starvation conditions may be due to the low rate of endogenous respiration by the bdellocysts. Bdellocysts are capable of germination in the presence or absence of prey cells in rich medium such as peptone-yeast extract.

Bacterial Proteins↗

Ultrastructural changes during encystment and germination of Bdellovibrio sp.

Under proper conditions, Bdellovibrio sp. strain W cells develop into bdellocysts in appropriate prey bacteria. After attachment and penetration of the prey cell, the encysting bdellovibrio began to accumulate inclusion material and increase in size, and was surrounded by an outer layer of amorphous electrondense material. The cytoplasm of the encysting cell appeared more electron dense, and nuclear areas appeared more compact. During germination of bdellocysts, the outer wall was uniformly broken down the inclusion material changed shape and affinity for the heavy metal stain, and the nuclear areas expanded. As the outer wall was dissolved, outgrowth began with the elongation of the germinant as it emerged from the prey ghost as an actively motile cell.

Bdellovibrio↗

Chemotaxis by Bdellovibrio bacteriovorus toward prey.

A chemotaxis assay system that uses a modified Boyden chamber was characterized and used for measurements of chemotaxis by Bdellovibrio bacteriovorus strain UKi2 toward several bacterial species. Bacteria tested included both susceptible and nonsusceptible cells (Escherichia coli, Pseudomonas fluorescens, Bacillus megaterium, and B. bacteriovorus strains UKi2 and D). None was attractive to bdellovibrios when present at densities below 10(7) cells per ml. Chemotaxis toward E. coli was studied most extensively; under conditions that minimized effects of osmotic shock to the cells, E. coli and exudates from E. coli at densities as high as 10(8) cells per ml failed to elicit a chemotactic response. Cell-free filtrates from mixed cultures of bdellovibrios and E. coli neither attracted nor repelled bdellovibrios. The data indicate that bdellovibrios do not use chemotaxis to locate prey cells.

Bacillus megaterium↗

Morphogenesis and ultrastructure of Geotrichum candidum septa.

The ultrastructure and mode of formation of septa of Geotrichum candidum were investigated by light and electron microscopy. The invaginations of the lateral membrane and wall appear to initiate at multiple points around the circumference of the cell; the immature septum subsequently assumes a cart-wheel shape, with branched spokes radiating from the center of the septum. Each face of the septum is covered with a membrane possessing hitherto undescribed structural differentiation; the membrane substructures are comprised of two central subunits encircled by 12 identical subunits. The diameter of the entire 12 plus 2 structure is 24 to 25 nm, and the diameter of each individual subunit is approximately 4 nm. The maturation of the septum appears to occur by further deposition of material along the branched skeletal regions. Numerous small openings (micropores), formed as a result of incomplete deposition, ultimately give rise to plasmodesmata. During arthrospore formation, the plasmodesmal canals and associated micropores are occluded by electron-dense materials, rendering each segment of the hyphae completely independent of the rest of the hyphae.

Cell Membrane↗

Occurrence of phosphonosphingolipids in Bdellovibrio bacteriovorus strain UKi2.

The major phospholipids of two strains of Bdellovibrio bacteriovorus were characterized. Both strain UKi1, which is obligately saprophytic, and strain UKi2, which is facultatively parasitic, contained phosphatidylethanolamine and phosphatidylglycerol as their major glycerophosphatides. A branched, 15-carbon fatty acid is the major component of these alkali-labile lipids. Absent from UKi1 but present in UKi2 were three alkali-stable lipids (compounds 8, 9, and 11) which appear to be phosphosphingolipids. After acid hydrolysis, both compound 8 and 9 yield the identical phosphorus-containing substance that is water soluble, dipolar ionic, and ninhydrin positive. This substance appears to contain a C-P bond since P(i) could not be released from this substance by treatment with alkaline phosphatase or by very harsh mineral acid treatment. Based on chromatographic comparisons, this phosphonate appears to be a novel lipid constituent. Upon degradation, compound 8 yields 1 mol of dihydroxy long-chain base and compound 9 yields 1 mol of a trihydroxy long-chain base. These bases appear to have a 17-carbon, possibly branched, structure based on gas-liquid chromatography retention times. Degradation of both sphingolipids yields a mixture of hydroxy fatty acids, the major component being a branched, 15-carbon hydroxy acid.

Alkaline Phosphatase↗

Morphogenesis of ascospores in Saccharomyces cerevisiae.

Ultrastructural changes associated with ascospore formation in Saccharomyces cerevisiae were investigated by using freeze-etching and thin-sectioning techniques. The first nuclear division (meiosis I) is indicated by the appearance of spindle fibers within the nucleus. The nucleus subsequently elongates and eventually assumes a barbell shape; the second nuclear division (meiosis II) occurs before nuclear separation. The spindle fibers involved in meiosis II appear to be oriented perpendicular to those observed in meiosis I. A discrete bilaminar structure (forespore wall) progressively delineates each ascospore nucleus and encloses cytoplasmic material including mitochondria and endoplasmic reticulum. The forespores then elongate, close off, and become separated from the ascus cytoplasm by membranes. The ascospores assume a spherical shape as spore coat material is laid down; the latter stages of ascospore formation are characterized by thickening of the ascospore wall and disintegration of the ascus cytoplasm. No structures which could be identified as chromosomes were observed.

Cell Nucleus↗

Isolation and preliminary characterization of bacteriophages for Bdellovibrio bacteriovorus.

Ten bacteriophages that attack and lyse saprophytic strains of Bdellovibrio bacteriovorus were isolated. Morphological, serological, and host-range studies revealed that there were four different bdellovibrio phages present among the isolates. One of the phages lysed a strain of B. bacteriovorus that requires the presence of a suitable bacterial host for growth. The phage attached to the bdellovibrio cells in the absence of the bacterial host cells; lysis occurred only in the presence of host cells. The 19 saprophytic bdellovibrio strains employed in the phage host-range studies were grouped on the basis of their susceptibility to phage lysis.

Antigens, Viral↗

Ultrastructural changes associated with activation and germination of Bacillus cereus T spores.

The ultrastructural changes occurring during defined stages of the transition of dormant Bacillus cereus T spores into heat-sensitive forms were investigated. The coat of the heat-activated spores displayed conspicuous striations across its middle layer. Electron microscopy of thin sections of heat-activated spores revealed the presence in the spore of a layer consisting of hexagonally oriented subunits. It was demonstrated that the subcoat region, but not the cortex, disappears rapidly during germination of B. cereus T spores. The fibrous structures apparently associated with the spore coat remain virtually unchanged during the entire course of activation and germination.

Alanine↗

Appendage development in Clostridium bifermentans.

The appendages of Clostridium bifermentans UK-A 1003 spores were shown to originate from a substance located just exterior to the outer forespore membrane. The dense spore coat develops along the periphery of this material, and, as the appendages develop in the cytoplasm, the coalescing spore coat intervenes between the appendages and their origin. Freeze etching revealed that the appendages are in the form of distinct fibers in proximity to the mature spore body. These fibers form a network around the spore, seemingly encasing it and insuring that the appendages remain attached to the mature, free spore. The inner wall of each appendage tubule is lined with fibers whereas the outer surface is smooth. The developing exosporium contained several layers consisting of small (3 nm) globular subunits; the outer exosporial surface is composed of relatively unstructured material.

Cell Membrane↗

A thermophilic, acidophilic mycoplasma isolated from a coal refuse pile.

A thermophilic, acidophilic procaryote lacking a cell wall has been isolated from a coal refuse pile which had undergone self-heating. Electron micrographs, chemical assays for hexosamine, and the inability of vancomycin to inhibit growth confirm the lack of a cell wall. The apparent ability of the organism to reproduce by budding and the low guanine plus cytosine content of its DNA indicate a relation to the mycoplasmas. The temperature optimum of the organism is 59 degrees C, and growth occurs over a range of 45 degrees to 62 degrees C. No growth occurs at 37 degrees C or at 65 degrees C. The optimum pH for growth is between 1 and 2, and growth occurs between pH 0.96 and 3.5 but does not occur at pH 0.35 and only poorly at pH 4.0. We propose to call this organism Thermoplasma acidophila. The existence of this organism extends considerably the range of habitats in which mycoplasma may occur.

Cell Wall↗

Ultrastructural changes associated with germination and outgrowth of an appendage-bearing clostridial spore.

The sequence of events occurring during the germination and outgrowth of appendage-bearing spores of Clostridium bifermentans was studied by phase-contrast and electron microscopy. The mature spore was characterized ultrastructurally as having the normal spore components as well as long tubular appendages which orginated from the surface of the spore coat. Spores were incompletely enclosed by a distinctly laminated exosporium which possessed hairlike projections on its outermost layer. During germination, structural changes were observed in the core, core wall, cortex, and spore coat layers. Cortical material was extruded from the spore during outgrowth, which usually occurred from the pole opposite the appendages. The subunits comprising the structure of the appendages and the morphology of the mature appendages were observed. No discernible changes could be observed in the spore appendages during germination and outgrowth.

Bacteriological Techniques↗

Facultatively parasitic strain of Bdellovibrio bacteriovorus.

A strain of Bdellovibrio bacteriovorus (designated strain UKi2) was isolated which was capable of growing either saprophytically in host-free medium or endoparasitically in Escherichia coli B/r. It was quantitatively determined that each bdellovibrio could develop in solid medium to produce a colony, and 65% of the cells in a late exponential-phase culture were capable of inducing E. coli B/r spheroplasts. A photomicrographic sequence of single E. coli spheroplasts containing bdellovibrios demonstrated that parasitically derived B. bacteriovorus UKi2 could develop saprophytically after release from the host cells. Strain UKi2 appears to be morphologically quite similar to previously described obligately parasitic bdellovibrios; biochemical data on this strain suggests its close relationship to some of the previously described host-independent strains of Bdellovibrio.

Bacteria↗

Ultrastructure and cell division of a facultatively parasitic strain of Bdellovibrio bacteriovorus.

Some aspects of cell development and division of Bdellovibrio bacteriovorus strain UKi2 were examined by use of electron microscopic techniques. Under saprophytic and parasitic conditions of growth, the comma-shaped cells enlarge, elongate, and form helical filaments. The mechanism of division appears to consist of an asymmetrical constriction of the filamentous cell by the cytoplasmic membrane, accompanied by a breakdown of the outer layers of the cell wall in the division region. During regeneration of the cell wall, the flagellum and flagellar sheath are formed. The development of the flagellum of the daughter cell is initiated prior to separation of the newly formed cells from the filament. Observations of B. bacteriovorus UKi2 grown under saprophytic and parasitic conditions indicate that development and ultrastructure are similar in both modes of growth.

Bacteria↗