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Casida, L. E., Jr.

Publications and source records attributed to Casida, L. E., Jr..

Minute tubular forms in soil

Large numbers of long, straight, flattened structures were observed during an electron-microscope study of bacteriophage in aqueous extracts. These structures were called tubules and ranged in width from 10 to 50 nm. Materials and methods were discussed relative to extraction of tubules from soil, electron microscopy, quantitation of tubules in filtrate suspension, tests performed on tubules, plaque formation, and nutrient amendments. It is found that all of the tubules recovered from soil are broken at one or both ends. They are present in surface soils but not in a subsurface sample. Their numbers decrease during bacterial multiplication in soil or broth-containing soil. The tubules appear to be composed of protein that could be disintegrated to liberate nonprotein fibers. A possible clue to the nature of the tubules is their apparent relation to soil bacteria.

Casida, L. E., Jr.

Survival and reversion of a stable L form in soil

The stable L form of Agromyces ramosus reverted to a bacterial form when incubated in sterilized soil. The cellular and colonial morphology of this bacterial form resembled that of the original parent bacterial form. The two forms differed, however, in that the revertant maintained its bacterial form when transferred onto a low-salt (NaCl) medium but was virtually completely induced into the L-form state on a high-salt medium. The original parent bacterial form was not sensitive to salt. The possibility is discussed that an L-form - bacterial-form cycle for this bacterium might occur naturally in soil. This cycle would be mediated by fluctuations in local salt concentrations in the soil.

Horwitz, A. H.

Microbial metabolic activity in soil as measured by dehydrogenase determinations

The dehydrogenase technique for measuring the metabolic activity of microorganisms in soil was modified to use a 6-h, 37 C incubation with either glucose or yeast extract as the electron-donating substrate. The rate of formazan production remained constant during this time interval, and cellular multiplication apparently did not occur. The technique was used to follow changes in the overall metabolic activities of microorganisms in soil undergoing incubation with a limiting concentration of added nutrient. The sequence of events was similar to that obtained by using the Warburg respirometer to measure O2 consumption. However, the major peaks of activity occurred earlier with the respirometer. This possibly is due to the lack of atmospheric CO2 during the O2 consumption measurements.

Casida, L. E., Jr.

New microbial growth factor

A screening procedure was used to isolate from soil a Penicillium sp., two bacterial isolates, and a Streptomyces sp. that produced a previously unknown microbial growth factor. This factor was an absolute growth requirement for three soil bacteria. The Penicillium sp. and one of the bacteria requiring the factor, an Arthrobacter sp., were selected for more extensive study concerning the production and characteristics of the growth factor. It did not seem to be related to the siderochromes. It was not present in soil extract, rumen fluid, or any other medium component tested. It appears to be a glycoprotein of high molecular weight and has high specific activity. When added to the diets for a meadow-vole mammalian test system, it caused an increased consumption of diet without a concurrent increase in rate of weight gain.

Bok, S. H.

Release of microorganisms from soil with respect to transmission electron microscopy viewing and plate counts

A study was conducted to obtain information concerning the fate of soil microorganisms during the procedures normally used in separating them from the soil in connection with various types of investigations. A silty clay loam (pH 6.0, moisture content 25%, organic content 3.5%) was used in the study. The results of the study indicate that many of the microbial cells naturally residing in soil remain attached to or, for some other reason, are not separated from the soil debris despite the use of various combinations of blending, sonication, and chemical dispersing agents. The method B used by Balkwill et al. (1975) provides a reasonable electron microscopy evaluation of the soil microflora.

Balkwill, D. L.

L-phase variants of Agromyces ramosus

Earlier results suggested that Agromyces ramosus possibly might exist naturally in soil as a cell-wall-defective form. The purpose of the present study was to test this hypothesis by determining whether the laboratory-adapted strains of A. ramosus could be artificially induced into the L-phase and, if so, to examine some parameters affecting induction and the stability of the L-forms. The hypothesis was also tested by attempting to revert the laboratory L-phase strains by subjecting them to the technique originally used for isolation of the bacterial form from soil. It is shown that A. ramosus is easily induced into the L-phase by growing it on an agar media containing low levels of penicillin or glycine. The L-forms are found to be stable after initial contact with the inducing agent and to be unable to be reverted to the bacterial form. However, this lack of reversion does not completely negate the hypothesis that L-forms might occur in nature, because it is possible that L-forms existing in the natural state are less stable than those found in the laboratory where there is little selective pressure toward reversion.

Horwitz, A. H.

Arthrobacter globiformis and its bacteriophage in soil

An attempt was made to correlate bacteriophages for Arthrobacter globiformis with soils containing that bacterium. The phages were not detected unless the soil was nutritionally amended (with glucose or sucrose) and incubated for several days. Phage was continuously produced after amendment without the addition of host Arthrobacter. These results indicate that the bacteriophage is present in a masked state and that the bacteria are present in an insensitive form which becomes sensitive after addition of nutrient.

Casida, L. E., Jr.

Plating isolation of various catalase-negative microorganisms from soil

A unique plating procedure was developed that allows isolation, but not enumeration, of representatives of the catalase-negative soil microflora. The numbers recovered, however, are low as compared to the numbers recovered when the modified dilution-to-extinction isolation procedure is used. The latter procedure provides prolonged inoculation in sealed tubes containing a nutritionally rich broth medium over small submerged agar slants. In contrast, the plating procedure utilizes nutritionally minimal media and the shorter incubations mandated by the inherent problems associated with plating.

Labeda, D. P.

Responses of indigenous microorganisms to soil incubation as viewed by transmission electron microscopy of cell thin sections.

Indigenous soil microorganisms were cultivated in their soil habitat with 50% moisture capacity at 30 C for two weeks. Changes in microorganism cells were studied by electron microscopy during incubation, with particular attention to the dormant cell growth and to the ability of cystlike cells to germinate and reencyst. The responses of various cell species to incubation conditions are described and illustrated by photomicrographs.

Bae, h. C.

Interval scanning photomicrography of microbial cell populations.

A single reproducible area of the preparation in a fixed focal plane is photographically scanned at intervals during incubation. The procedure can be used for evaluating the aerobic or anaerobic growth of many microbial cells simultaneously within a population. In addition, the microscope is not restricted to the viewing of any one microculture preparation, since the slide cultures are incubated separately from the microscope.

Casida, L. E., Jr.