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R M Pfister

Publications and source records attributed to R M Pfister.

At least 37 records · Page 2Linked to original sources

Improved staining of extracellular polymer for electron microscopy: examination of Azotobacter, Zoogloea, Leuconostoc, and Bacillus.

Phase contrast, ultraviolet microscopy, and freeze-etching were used to determine the amount of exocellular polymer surrounding unfixed cells of four genera of bacteria: Azotobacter vinelandii, Zoogloea ramigera, Leuconostoc mesenteroides, and an acid-tolerant, floc-forming Bacillus species. Thin-sectional electron microscopy was employed to measure the effectiveness of a modified ruthenium red staining method. The results obtained with this modification of ruthenium red staining technique were compared to results obtained when previously proposed ruthenium red methods of fixation were employed. The results of these relations were then compared to the amounts of exocellular material as determined with phase-contrast microscopy, ultraviolet microscopy, and freeze-etching. The data obtained indicate that improved fixation of exocellular polymer is achieved when cells are pretreated with ruthenium red as described herein. In addition, the modified methods also reveal cytological detail not apparent when other methods of ruthenium fixation are employed.

Azotobacter↗

Freeze-etching of Azotobacter vinelandii: examination of wall, exine, and vesicles.

The structure of the cell wall, the arrangement of the cyst exine, and the origin and distribution of intine vesicles in Azotobacter vinelandii ATCC 12837 were examined by freeze-etching and conventional electron microscopic techniques. In the vegetative organism the cell wall appears to have a woven texture which disappears during cyst formation. The exine is composed of two different types of material: the outer layer is a fibrous, amorphous layer, and the numerous inner layers form the basic hexagonal structures which unite to form the cyst coat. The presence of intine vesicles in the encysting organism was confirmed in frozen-etched cells. The appearance of frozen-etched cells and cysts and the distribution of capsular material indicate that extracellular polysaccharide is an important factor in cyst formation.

Azotobacter↗

Aldrin: removal from lake water by flocculent bacteria.

Floc-forming bacteria isolated from Lake Erie adsorb and concentrate aldrin from colloidal dispersion so that the settling of the bacterial flocs removes aldrin from the water phase. Contemporary sediments forming in Lake Erie contain aldrin and could adsorb more. The sediments consist of a conglomerate floc of bacteria, diatoms, and inorganic and detrital particles. Flocculent bacteria also adsorb microparticulates, and this adsorption capacity represents a mechanism for sediment formation and for the removal of suspended particles including aldrin from the water column.

Adsorption↗

Biological nitrogen fixation in Lake Erie.

Biological nitrogen fixation, as determined by acetylene reduction, occurs in Lake Erie. Fixation potential by blue-green algae in situ in water and by bacteria in collected sediments was demonstrated. Nitrogen-fixing activity occurred from June through November suggesting that it is significant over the extremes of seasonal variation in light, temperature, and nutrients.

Acetylene↗

Aerobic heterotrophic bacteria indigenous to pH 2.8 acid mine water: microscopic examination of acid streamers.

"Acid streamers" found in acid coal mine drainage consist of bacteria trapped within an extracellular fibrillar polymer network. Inorganic compounds also precipitate within the polymer network. Several bacteria which appear to be different and are presumed to be different species are associated in the slimy mass of the "acid streamers." The "streamers" contain individual microcolonies or microcosms that can be recognized by a selective polysaccharide stain, which suggests that the slime streamer is a conglomeration of polymers produced by more than one species.

Bacteria↗

Aerobic heterotrophic bacteria indigenous to pH 2.8 acid mine water: predominant slime-producing bacteria in acid streamers.

Five gram-negative bacteria, two gram-positive bacteria, and one yeast were isolated from "acid streamers" taken from acid mine water. One gram-positive rod which has been tentatively identified as a Bacillus species appeared to be the predominant organism in the streamers. This isolate produced copious amounts of extracellular polymer at 10 C in the laboratory and was considered to be the primary source of polymer in the "acid streamer" slime matrix. The organism grew slowly at pH 2.8 in mine water media, but the optimal pH was approximately 7.0.

Bacillus↗

Microparticulates: isolation from water and identification of associated chlorinated pesticides.

Microparticulates suspended in lake water were collected by continuous centrifugation and either examined directly or placed on a linear sucrose gradient. Total residue as well as fractions of the centrifuged gradient were extracted with hexane and examined by gas chromatography for the presence of chlorinated hydrocarbon pesticides. Hexane extracts of total residues were also examined by thin-layer chromatography. Lindane and endrin were shown, by gas-liquid chromatography and thin-layer chromatography, to be associated with microparticles. These and other pesticides appeared to be selectively associated with microparticles of different densities, when gas-liquid chromatography was used, although concentrations were below the detection limits required for confirmation by thin-layer chromatography. Samples taken at different times from different locations in Lake Erie revealed different associations with hexane-soluble electron-capturing compounds.

Centrifugation, Density Gradient↗

Structure of exocellular polymers and their relationship to bacterial flocculation.

Several gram-negative, polarly flagellated rods were isolated on the basis of their flocculent growth habit. Some of the isolates possessed a capsular matrix which is composed of exocellular fibrils. Other isolates did not appear to have a capsular matrix when examined with a bright-field microscope with or without the aid of stains. However, these latter type isolates did possess exocellular material which can be demonstrated by adsorption of a fluorescent dye under an ultraviolet microscope. Electron microscopic examination demonstrated that the exocellular material around all isolates examined is fibrillar. The fibrils were susceptible to cellulase although all fibrils did not appear to be identical. It is postulated that the exocellular polymers were responsible for the flocculent growth habit of the bacteria, and that the process of bacterial flocculation produced by synthetic polyelectrolytes was essentially the same as that caused by naturally produced exopolymers.

Bacteria↗