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R A Neihof

Publications and source records attributed to R A Neihof.

8 recordsLinked to original sources
PubMed1984

Direct determination of activities for microorganisms of chesapeake bay populations.

We used three methods in determination of the metabolically active individual microorganisms for Chesapeake Bay surface and near-bottom populations over a period of a year. Synthetically active bacteria were recognized as enlarged cells in samples amended with nalidixic acid and yeast extract and incubated for 6 h. Microorganisms with active electron transport systems were identified by the reduction of a tetrazolium salt electron acceptor. Microorganisms active in uptake of amino acids, thymidine, and acetate were determined by microautoradiography. In conjunction with enumeration of active organisms, a total direct count was made for each sample preparation by epifluorescence microscopy. For the majority of samples, numbers of amino acid uptake-active organisms were greater than numbers of organisms determined to be active by other direct measurements. Within a sample, the numbers of uptake-active organisms (amino acids or thymidine) and electron transport system-active organisms were significantly different for 68% of the samples. Numbers of synthetically active bacteria were generally less than numbers determined by the other direct activity measurements. The distribution of total counts in the 11 samplings showed a seasonal pattern, with significant dependence on in situ water temperature, increasing from March to September and then decreasing through February. Synthetically active bacteria and amino acid uptake-active organisms showed a significant dependence on in situ temperature, independent of the function of temperature on total counts. Numbers of active organisms determined by at least one of the methods used exceeded 25% of the total population of all samplings, and from June through September, >85% of the total population was found to be active by at least one direct activity measurement. Thus, active rather than dormant organisms compose a major portion of the microbial population in this region of Chesapeake Bay.

Journal Article↗
PubMed1983

Inhibitory effect of solar radiation on amino Acid uptake in chesapeake bay bacteria.

The effect of solar radiation on a natural bacterial population from the Chesapeake Bay was evaluated from measured changes in numbers of organisms engaged in amino acid uptake. From July through May, freshly collected water samples were exposed in quartz containers to 3.5 h of total sunlight both with and without UV-absorbing filters. Water samples were subsequently incubated with tritiated amino acids, and the uptake-active bacteria were assayed by microauto-radiography-epifluorescence microscopy. The survival index, defined as the fraction of the uptake-active population that remained active after the exposure to sunlight, ranged from 0.93 to 0.20. Decreased survival was correlated with increased solar intensity. The inhibition of amino acid uptake was attributed not only to the UV-B component of the solar spectrum (280 to 320 nm), but also to longer UV and visible wavelengths.

Journal Article↗
PubMed1982

Improved method for determination of respiring individual microorganisms in natural waters.

A method is reported that combines the microscopic determinations of specific, individual, respiring microorganisms by the detection of electron transport system activity and the total number of organisms of an estuarine population by epifluorescence microscopy. An active cellular electron transport system specifically reduces 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride (INT) to INT-formazan, which is recognized as opaque intracellular deposits in microorganisms stained with acridine orange. In a comparison of previously described sample preparation techniques, a loss of >70% of the counts of INT-reducing microorganisms was shown to be due to the dissolution of INT-formazan deposits by immersion oil (used in microscopy). In addition, significantly fewer fluorescing microorganisms and INT-formazan deposits, both </=0.2 mum in size, were found for sample preparations that included a Nuclepore filter. Visual clarity was enhanced, and significantly greater direct counts and counts of INT-reducing microorganisms were recognized by transferring microorganisms from a filter to a gelatin film on a cover glass, followed by coating the sample with additional gelatin to produce a transparent matrix. With this method, the number of INT-reducing microorganisms determined for a Chesapeake Bay water sample was 2-to 10-fold greater than the number of respiring organisms reported previously for marine or freshwater samples. INT-reducing microorganisms constituted 61% of the total direct counts determined for a Chesapeake Bay water sample. This is the highest percentage of metabolically active microorganisms of any aquatic population reported using a method which determines both total counts and specific activity.

Journal Article↗
PubMed1982

Improved microautoradiographic method to determine individual microorganisms active in substrate uptake in natural waters.

We report a method which combines epifluorescence microscopy and microautoradiography to determine both the total number of microorganisms in natural water populations and those individual organisms active in the uptake of specific substrates. After incubation with H-labeled substrate, the sample is filtered and, while still on the filter, mounted directly in a film of autoradiographic emulsion on a microscope slide. The microautoradiogram is processed and stained with acridine orange, and, subsequently, the filter is removed before microscopic observation. This novel preparation resulted in increased accuracy in direct counts made from the autoradiogram, improved sensitivity in the recognition of uptake-active (H-labeled) organisms, and enumeration of a significantly greater number of labeled organisms compared with corresponding samples prepared by a previously reported method.

Journal Article↗
PubMed1978

Biocidal properties of anti-icing additives for aircraft fuels.

The biocidal and biostatic activities of seven glycol monoalkyl ether compounds were evaluated as part of an effort to find an improved anti-icing additive for jet aircraft fuel. Typical fuel contaminants, Cladosporium resinae, Gliomastix sp., Candida sp., Pseudomonas aeruginosa, and a mixed culture containing sulfate-reducing bacteria were used as assay organisms. Studies were carried out over 3 to 4 months in two-phase systems containing jet fuel and aqueous media. Diethylene glycol monomethyl ether, triethylene glycol monomethyl ether, triethylene glycol monoethyl ether, and 2-methoxyethanol were generally biocidal in aqueous concentrations of 10 to 17% for all organisms except Gliomastix, which required 25% or more. 2-Ethoxyethanol, 2-propoxyethanol, and 2-butoxyethanol were biocidal at progressively lower concentrations down to 1 to 2% for 2-butoxyethanol. The enhanced antimicrobial activity of these three compounds was attributed to cytoplasmic membrane damage because of the correlation between surface tension measurements and lytic activity with P. aeruginosa cells. The mechanism of action of the less active compounds appeared to be due to osmotic (dehydrating) effects. When all requirements are taken into account, diethylene glycol monomethyl ether appears to be the most promising replacement for the currently used additive, 2-methoxyethanol.

Aerobiosis↗
PubMed1978

Chemical modification of the surfaces of bacterial cell walls.

The surfaces of the isolated cell walls of four bacterial species were studied by microelectrophoresis following chemical treatments intended to remove specific charged groups. Acid-base titrations of the walls were used to assess specificity and extent of the modifications. Carboxyl groups were specifically and completely modified by activation with a water-soluble carbodiimide and subsequent reaction with a nucleophile, such as glycinamide, to give an uncharged pH-stable product. Aqueous media and mild reaction conditions make the method suitable for modifying carboxyl groups on cell surfaces too labile to withstand the harsh conditions required for conventional esterification reactions. Use of the carbodiimide-mediated reaction for discharging carboxyl groups, along with fluorodinitrobenzene for discharging amino groups and extraction procedures for removing constituents carrying phosphoester groups (teichoic acids), made it possible to obtain information about the spatial arrangement of charged groups on the wall surfaces. Removal of the exterior negative charge dominating wall surfaces allowed underlying amino groups to become electrokinetically effective and, in the case of E. coli, also revealed a lipophilic region with an affinity for a cationic surfactant.

Bacillus↗