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Biomedical subjects

K J Cheng

Publications and source records attributed to K J Cheng.

At least 109 records · Page 6Linked to original sources

Maceration of Clover and Grass Leaves by Lachnospira multiparus.

A strain of Lachnospira multiparus, a pectin-hydrolyzing bacterium from the rumen, was incubated in nutrient media in the presence of surface-disinfected clover leaflets. When the culture flasks containing the leaflets together with Lachnospira were shaken after overnight incubation, extensive maceration of the leaflets was seen, although uninoculated control leaflets remained intact during a similar treatment. Examination of inoculated leaflets by transmission electron microscopy showed extensive invasion of intercellular areas of the mesophyll tissue but only minor invasion of vascular tissue. Cutting the leaves before incubation greatly increased the ability of L. multiparus to colonize and macerate the leaflets. Similar experiments with grass leaves are also described, and the possible role of maceration in the digestion of plant material in the rumen is discussed. Although Lachnospira stains gram variable and often gram negative, the ultrastructure of the cell wall was that of a gram-positive bacterium.

Journal Article↗

Production of alkaline phosphatase by epithelial cells and adherent bacteria of the bovine rumen and abomasum.

Three distinct bacterial populations have been defined in the bovine rumen: the rumen fluid population; the population associated with food particles; and the population adherent to the rumen epithelium. Alkaline phosphatase activity has been reported in cells of the first two populations and here we report that assays of rumen epithelial samples containing both tissue and bacteria also contain the enzyme. The reaction product technique has localized the enzyme both in adherent bacteria and in cell of the stratified squamous rumen epithelium. The epithelium of the abomasum shows much lower levels of alkaline phosphatase activity.

Abomasum↗

Adhesion of bacteria to epithelial cell surfaces within the reticulo-rumen of cattle.

Blocks of tissue were removed from various locations in the bovine digestive tract and fixed and processed for transmission and scanning electron microscopy by techniques that retained adherent bacteria. The distribution of bacteria on the surface of epithelial cells was examined by scanning electron microscopy. This showed intermittent colonization of the epithelia with the formation of occasional microcolonies of morphologically similar bacterial cells. Transmission electron microscopy of ruthenium red-stained material showed the presence of both the glycocalyx of the bovine epithelial cells and fibrous carbohydrate coats surrounding adherent bacteria. The carbohydrate coats appeared to mediate the attachment of bacteria to the epithelium, to food particles, and to each other so that microcolonies were formed. Careful examination of the bacterial colonization of keratinized cells in the process of being sloughed from the surface of the stratified squamous epithelium of the rumen showed that these dead cells were digested by adherent bacteria of a limited number of morphological types. The spatial relationship of this mixed, adherent, microbial population to living and dead epithelial cells and to food particles indicates that digestive processes of some importance may be accomplished by this stationary component of the microbial flora of the digestive tract.

Animals↗

Characterization of a Cytoplasmic Reserve Glucan from Ruminococcus albus.

Ruminococcus albus, an anaerobic bacterium that digests cellulose in the rumen of cattle, produces intracellular polysaccharide granules varying from 0.05 to 0.31 mum in diameter when grown in batch culture. This polysaccharide material was purified and found to contain d-glucose as the only reducing sugar. The polyglucose polymer was slightly opalescent in aqueous solution and formed a strong reddish purple iodine complex with a maximum absorbance at 550 nm. Its infrared spectrum had characteristic absorption bands at 8.70, 9.25, and 9.75 mum and was identical with that of the amylopectin-glycogen type of Megasphaera elsdenii and that of the glycogen of enteric bacteria and beef liver. It reacted strongly with concanavalin A. Methylation analysis showed that the glucan contained 2,3,4,6-tetra-O-MeG-2,3,6-Tri-O-MeG-2,3-Di-O-MeG in a ratio of 8:84:8. Characterization of the products obtained by treatment with isoamylase indicates that the glucan of R. albus is of the glycogen type.

Journal Article↗

Alkaline phosphatase activity of rumen bacteria.

Of the 54 strains of rumen bacteria examined for alkaline phosphatase (APase) production, 9 of 33 gram-negative strains and none of 21 gram-positive strains produced the enzyme. The APase of the cells of the three strains of Bacteroides ruminicola that produced significant amounts of the enzyme was located in the periplasmic area of the cell envelope, whereas the enzyme was located in the strains of Selenomonas ruminantium and Succinivibrio dextrinosolvens was associated with the outer membrane. The localization of APase production in the cells of natural populations of rumen bacteria from hay-fed sheep was accomplished by reaction product deposition, and both the proportion of APase-producing bacteria and the location of the enzyme in the cell envelope of the producing cells could be determined. We suggest that this procedure is useful in detecting shifts in the bacterial population and the release of cell-bound APase that accompany feedlot bloat and other sequelae of dietary manipulation in ruminants.

Alkaline Phosphatase↗

Ultrastructure of Butyrivibrio fibrisolvens: a gram-positive bacterium.

The cells of bacteria of the genus Butyrivibrio are universally described as being gram negative, and they produce an unequivocal gram-negative reaction in the standard staining procedure. However, their cell walls contain derivatives of teichoic acid, which are characteristic of gram-positive cells. In this study, the cell walls of two representative strains of Butyrivibrio were of the gram-positive morphological type, as seen by electron microscopy, but they were very thin (12 to 18 nm). The thinness of these cell walls may account for the tendency of these cells to stain gram negatively in the standard staining procedure. Ruthenium red staining revealed an extracellular structure surrounding cells of Butyrivibio sp. (strain C3). This structure was composed of individual "knobs" that sometimes mediated cell-to-cell adhesion in the culture.

Cell Membrane↗

Rumen bacteria: interaction with particulate dietary components and response to dietary variation.

The bovine rumen resembles many other ecosystems in that its component bacterial cells are universally surrounded and protected by extracellular structures. The most common form of these structures is a fibrous carbohydrate slime that extends away from the cell and may mediate the attachment of the bacterium to a surface. This attachment is relatively specific and it may occur at the surface of the rumen epithelium or on the cell walls of a specific tissue within the plant-derived food of the animal. The production of the extracellular slime is under nutritional control and slime may be overproduced when soluble carbohydrates are available in high concentration. This overproduction results in cell-cell adhesion among the rumen bacteria with the eventual formation of slime-enclosed microcolonies and, in extreme cases, the generation of sufficient viscosity to cause feedlot bloat.

Animal Feed↗

Frothy feedlot bloat in cattle: production of extracellular polysaccharides and development of viscosity in cultures of Streptococcus bovis.

Streptococcus bovis was cultured in a synthetic medium with three concentrations of sucrose. Initial viscosity of the media was 1.5 centipoise (cp). After incubation for 8 h, the viscosity of the medium with 0.5% sucrose was unchanged, that with 3% sucrose had increased to 8 cp, and that with 6% sucrose to 112 cp. Similar results were found with a rumen fluid medium. A slimy material, responsible for increased viscosity of these cultures, was digested by dextranase. The material appeared as a complex system of intercellular fibers when viewed under the electron microscope after freeze-etching. With proteins and other polymers released from lysed bacteria, this slimy material may contribute directly to increased viscosity and foam formation. In addition to these intercellular fibers, each cell was surrounded by a fibrous capsule that was not digested by dextranase. This capsule stained with lead citrate and uranyl acetate, but not with ruthenium red. The amount of capsular material produced was similar whether the media contained 0.5, 3.0, or 6% sucrose.

Anaerobiosis↗

Release of bacterial alkaline phosphatase in the rumen of cattle fed a feedlot bloat-provoking diet or a hay diet.

Alkaline phosphatase (APase) was present in the bovine rumen in both cell-free and cell-associated states and levels of the enzyme varied with dietary regime. Reaction product deposition showed that the enzyme was associated with the mixed bacterial population. No enzyme was observed to be associated with protozoa. Trace activity of APase was also detected in the saliva. The presence of large amounts of APase in cell-free rumen fluid of cattle fed fine concentrate feed is believed to be due, in part, to the breakage of bacterial cells that occurs in the rumen.

Alkaline Phosphatase↗

Ultrastructure of cell envelopes of bacteria of the bovine rumen.

Most of the bacteria found in rumen fluid samples taken from cows fed hay, or a concentrate diet, had cell walls of the gram-negative type. Most were intact, with only a small proportion of lysed cells, and many of the cells contained electron-translucent cytoplasmic deposits similar to the carbohydrate reserve material described in pure cultures of rumen organisms. All of the bacteria observed in these samples had an external "coat" layer outside the outer membrane when fixed in glutaraldehyde and osmium, stained with uranyl acetate and lead citrate, and examined as sectioned material. These coat layers varied from thin (ca. 8 nm) structures to very extensive fibrous systems, sometimes including concentric arrangements and radial fibers extending up to 1,200 nm from the cell. The thin-coat layers sometimes exhibited a rough periodicity. In all, 10 different types of coat layers were distinguishable on a morphological basis. It is proposed that these external coat layers have protective and adherence functions for the rumen bacteria in the environment.

Animals↗

Ultrastructure and adhesion properties of Ruminococcus albus.

Morphological studies have shown that cells of the anaerobic rumen bacterium Ruminococcus albus have electron-translucent granules of reserve carbohydrate in their cytoplasm, and that they have a polysaccharide "coat" layer external to their gram-negative cell wall. This coat layer, which stains specifically with ruthenium red, forms a compact mat of fibers adjacent to the cell, and fibrous elements also project as much as 0.6 mum from the cells. These radial fibers are clearly visualized by freeze-etching, and can be seen to extend throughout the extensive intercullular space in centrifuged pellets of these bacteria. Cells of R. albus adhere to cellulose fibers added to the culture medium, and the coat material is seen to mediate this adhesion in addition to its function in the general protection of these cells.

Anaerobiosis↗

Cell envelope morphology of rumen bacteria.

The cell walls of three species of rumen bacteria (Bacteroides ruminicola, Bacteroides succinogenes, and Megasphaera elsdenii) were studied by a variety of morphological methods. Although all the cells studied were gram-negative and had typical cytoplasmic membranes and outer membranes, great variation was observed in the thickness of their peptidoglycan layers. Megasphaera elsdenii evidenced a phenomenally thick peptidoglycan layer whose participation in septum formation was very clearly seen. All species studied have cell wall "coats" external to the outer membrane. The coat of Bacteroides ruminicola is composed of large (approximately 20 nm) globules that resemble the protein coats of other organisms, whereas the coat of Bacteroides succinogenes is a thin and irregular carbohydrate coat structure. Megasphaera elsdenii displays a very thick fibrillar carbohydrate coat that varies in thickness with the age of the cells. Because of the universality of extracellular coats among rumen bacteria we conclude that the production of these structures is a protective adaptation to life in this particular, highly competitive, environment.

Adaptation, Biological↗