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Biomedical subjects

C I Mayfield

Publications and source records attributed to C I Mayfield.

17 recordsLinked to original sources

Transfer and expression of mesophilic plasmid-mediated degradative capacity in a psychrotrophic bacterium.

A psychrotrophic bacterium, originally isolated from a natural aquatic environment, was characterized and identified as Pseudomonas putida Q5 for use as a representative recipient for biodegradative genes from a mesophilic microorganism. The TOL plasmid pWWO of the mesophile P. putida PaW1 was successfully transferred by conjugation to the naturally isolated psychrotroph P. putida Q5, as shown by plasmid analysis by agarose gel electrophoresis. Expression of the genes encoded by the mesophilic TOL plasmid in the psychrotroph was shown by the fact that the transconjugant (designated P. putida Q5T) had the capacity to degrade and utilize toluate (1,000 mg/liter) as a sole source of carbon at temperatures as low as 0 degrees C. Comparison of growth rates over a wide temperature range (0 to 30 degrees C) indicated that the physiological activity of the transconjugant was not reduced and that the plasmid DNA from the mesophile and its encoded enzymes functioned effectively in the psychrotroph at temperatures well below those at which the mesophile could grow. The production and demonstrated functioning of P. putida Q5T illustrates the possibility of developing specific degradative capacities in bacteria which can readily function at low temperatures in chemically contaminated environments or in industrial wastewater treatment systems.

Benzoates

Effect of sequence of exposure to chlorophenols in short-term bacterial bioassays.

A bioassay using Pseudomonas fluorescens was affected by the sequence of exposure to pentachlorophenol (PCP) and 2,3,4,5-tetrachlorophenol (TCP). Surviving cells from standardized cell suspensions initially treated with PCP at concentrations ranging from 10 to 75 micrograms/ml, followed by removal of the toxicant, were not affected by a second dose of PCP at the same concentration. However, if the second dose was TCP, the test organism was sensitive to the second exposure. The most toxic sequence was an initial exposure to TCP followed by a second exposure to PCP. The response of the test organism to PCP and TCP was clearly dependent upon both the toxicant concentrations used and the sequence of toxicant addition.

Bacteria

Perturbation of lipid membranes by organic pollutants.

The ability of a range of organic pollutants--hexachlorobenzene, mirex(1,1a,2,2,3,3a,4,5,5,5a,5b,6-dodecachlorooctahydro-1,3,4-metheno-1H cyclobuta(cd) pentalene), 1,3,5-trichlorobenzene, 2,4,6-trichlorophenol, p-nitrophenol, p-chlorophenol, DDT, and pentachlorophenol--to perturb liposomes of dipalmitoyl phosphatidylcholine (DPPC) has been measured by differential scanning calorimetry. The degree of perturbation was measured by the increase in breadth of the main DPPC phase transition in both heating and cooling scans. DDT and the phenol derivatives were effective perturbers of phospholipid, broadening the transition by as much as 12-fold. Hexachlorobenzene and mirex did not perturb at all when mixed with DPPC at concentrations as high as 20 mol%, although 1,3,5-trichlorobenzene caused slight broadening of the main transition at this concentration. Perturbation is facilitated by the presence of a hydroxyl group on the benzene ring and hindered by increasing degrees of chloride substitution. An apparent correlation exists between the extent of phospholipid perturbation measured by differential scanning calorimetry and LD50 values for these compounds taken from the literature. This suggests the possibility of formulating an "index of perturbation" which could be used to screen certain classes of organic compounds for potential biological toxicity on a routine basis.

Animals

Nitrogen fixation and methane metabolism in a stream sediment-water system amended with leaf material.

The reduction of acetylene and the production and oxidation of methane in a stream sediment--water system amended with either fresh leaves or autumn-shed leaves in the presence and absence of air were studied. Net methane production by the sediment--water system occurred only when leaf material was added, with fresh leaves giving 2.2 times the methane accumulation as autumn-shed leaves. Static incubation in the presence of air had little effect on net methane production, with such production being about the same as (with fresh leaves) or 79% of (with autumn-shed leaves) the anaerobic rates. Acetylene reduction was more affected by the presence of air, with anaerobic rates being at very low levels. The presence of fresh leaves was again stimulatory, with the acetylene reduction rate being 5.5-fold greater than that occurring with autumn-shed leaves.

Air

A method for the detection and analysis of growth patterns of microorganisms in soil.

A fluorescence-staining technique using the magnesium salt of 8-anilino-1-naphthalene sulfonic acid is described and used to follow the changes in the distribution patterns of microorganisms in soils. A statistical procedure was used to determine the occurrence of significant differences in clumping of bacteria (i.e., production of colonies) in different regions of artificial soil-aggregate systems treated with nutrient solutions and also with a herbicide, Linuron. The response of soil microorganisms to glucose amendment was most marked in the aerobic, outer zone of aggregates. Linuron inhibited colony formation in aggregates treated with the herbicide. The method allows continued observations to be made on the same soil sample at intervals during incubation and os can be used to determine growth rates, inhibitory effects of chemicals, distribution patterns in soils, effects of added nutrients, and other effects where growth in situ is important.

Anilino Naphthalenesulfonates

Effect of naturally occurring apatites on growth and morphology of algae.

Crystals (30--100 micrometer) of selected naturally occurring apatite (Ca10 (PO4)6(OH, F)2) samples were added to P-free (less than 0.001 microgram/ml total P) Bristol's medium (1-1000 microgram/ml of apatite) as the sole source of ortho-PO43-. The media were inoculated with washed, non-axenic cells of three chlorophycean algal species cultivated under PO43--deficient conditions. Phase-contrast and scanning electron microscopy revealed that at low slurry densities (1-10 microgram/ml of apatite), Ankistrodesmus braunii (ATCC 2744) cells were morphologically distorted. At concentrations of 100 and 1000 microgram/ml of apatite, more than 85% of the cells had undergone autospore formation within 7--10 days of incubation at 20 degrees C. Most autospores formed failed to germinate under high nutrient conditions. Scenedesmus longus (No. 1236) formed colonies when cultivated in Bristol's medium but daughter cells displayed a Chodatella-like unicellular morphology when grown in apatite media. Test algal species (Chlamydomonas dysosmos, S. longus, A. braunii) showed a marked preference for growth on apatite crystals over non-nutritive surfaces. Unialgal and mixed-algal cultures produced an extensive matrix of extracellular fibrous material in response to growth on crystals at concentrations greater than 10 microgram/ml of apatite.

Apatites

A rapid, simple method for staining bacterial flagella.

A simple modification of Gray's flagellar staining procedure is described. It can be used on air-dried smears or directly on wet mounts of motile bacteria. The stained bacterial flagella can be observed with phase-contrast or bright-field optics. No rigorous cleaning of slides, counterstains, or any washing procedures are required with the staining method, making it very suitable for routine examinations.

Bacteria

A fluorescence staining method for the examination of microorganisms on natural substrates.

A fluorescence staining technique is described which uses the magnesium salt of 1-anilino-8-naphthalene sulfonic acid as a protein stain. At a concentration of 2.5 or 3.0 mg ml-1 in distilled water or phosphate buffer the compound does not fluoresce under ultra-violet or far blue illumination until it is bound to a protein or similar compound. It can be applied to natural substrates such as soil, food materials, organic material in water, etc., and can be examined immediately without any washing procedures. The application of this staining method to detect microorganisms on dead green algae (detritus) is described.

Anilino Naphthalenesulfonates

A simple fluorescence of staining technique for in situ soil microorganisms.

A simple, rapid straining technique using the magnesium salt of 1-anilino-8-naphthalene sulfonic acid is described. Treatment of soil with an aqueous, membrane-filtered solution (3.5 mg/ml) of the salt causes the soil microorganisms to fluoresce when examined with light from a mercury arc light source.

Anilino Naphthalenesulfonates