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A method for the quantitative determination of gibberellic acid in fermentation media, using descending paper chromatography in butylacetate-water is described. A sample is adjusted to pH 2.5–3.0, extracted with n-butanol, and 0.05 ml. organic layer is spotted on Chromatographic paper. After equilibration of the atmosphere in the jar, the chromatogram is developed in butylacetate saturated with water, for 7 hours, and the solvent is allowed to drip off the bottom of the sheet. Detection is carried out by spraying with a 3% solution of sulphuric acid in methanol and after drying the paper, spots with blue u.v. fluorescence are observed. The area of the spots is evaluated with the aid of a calibration curve, plotted from values of standards corresponding to 20, 60 and 120 μg. gibberellic acid. The error of estimation is ±10–15% when estimations are carried out carefully. The lowest limit of sensitivity is 5 μg.

SUMMARY: A simple method of detecting arabinose by paper chromatography in the cell-walls of aerobic actinomycetes is described. Mycobacteria and Nocardia species are rich in arabinose while saprophytic species of Streptomyces are deficient. Pathogenic Streptomyces species tend to fall between the two extremes but Streptomyces somaliensis is apparently nearly devoid of arabinose.

High rates of acetylene (C 2 H 2 ) reduction (nitrogenase activity) were observed in woodroom effluent from a neutral sulfite semi-chemical mill under aerobic (up to 644 nmol of C 2 H 4 produced per ml per h) and under anaerobic (up to 135 nmol of C 2 H 4 produced per ml per h) conditions. Pasteurized effluent developed C 2 H 2 reduction activity when incubated under anaerobic but not under aerobic conditions. Activities were increased by addition of 0.5 to 3.0% glucose or xylose. Enrichment and enumeration studies showed that N 2 -fixing Azotobacter and Klebsiella were abundant, and N 2 -fixing Bacillus was present. Of 129 isolates of Klebsiella from pulp mills, lakes, rivers, and drainage and sewage systems, 32% possessed nitrogen-fixing ability.

A convenient, sensitive, and reliable assay for the conversion of radiolabeled sucrose to alcohol-insoluble dextran by the Streptococcus mutans dextransucrase has been developed.

SUMMARY: The amino acid and peptide content of the micrococci as revealed by paper chromatography depended on the physiological age of the culture and on the composition of the growth medium. The use of different peptones in a Yeastrel glucose peptone medium resulted in minor changes in the chromatograms. A greater effect was noted when glucose was omitted. When the composition of the growth medium was standardized the best comparison of cultures was afforded by the preparation of chromatograms from samples taken in the late logarithmic and early stationary phases of growth.

Studies of serum glycoproteins by paper electrophoresis in rheumatic fever showed the following: Total glycoproteins were generally increased in concentration, the alpha-2 glycoprotein fraction showed moderate to marked increases in value, and alpha-1 and beta glycoprotein components generally showed moderately decreased concentrations.

SUMMARY A preliminary survey of species of the genus Bacillus has shown that the comparative study of the bacterial amino pool, using paper partition chromatography techniques, shows promise as an aid to differentiation. But special factors, such as sporulation and other growth characteristics, make the use of these techniques more complicated than is the case with some other genera. Seventy-five mesophilic strains were examined and in general the chromatogram patterns correlated well with the biochemical, morphological and cultural characters. Further, it appears that these chromatographic techniques could be applied with advantage in the study of spore germination, heat activation and sporulation, and of the influence of growth medium, pH, temperature and degree of aeration on the metabolism of micro-organisms. The taxonomy of the thermophilic sporeformers and of new species would be clarified by chromatographic studies.

The author describes a method, for the determination of bacterial L-histidine, L-tyrosine, L-tryptophan, L-arginine, L-lysine, L-ornithine, L-serine, L-phenyl-alanine, L-glutamic acid and L-aspartic acid decarboxylase by means of high voltage paper electrophoresis, using electrolytes of low ionic strength. For the group of test microorganisms (Enterobacteriaceae, Achromobacteriaceae andPseudomonas), L-histidine, L-arginine, L-lysine, L-ornithine and L-glutamic acid decarboxylases were found to be of the greatest practical significance. The optimal conditions for maximum formation of these decarboxylases by some of the test strains ofEscherichia coli, Hafnia andProteus are discussed.

SUMMARY: The use of electronic computation in the objective analysis of the chromato-graphic patterns of 90 strains of heterofermentative lactobacilli is described. The results showed that there was such a spread of properties among the different strains that a division into species was not justifiable.

SUMMARY: Chromatograms of unhydrolyzed extracts of the type strains of the Cowan groups of the micrococci and of other representatives of these groups have shown that the resulting pattern of ninhydrin-positive spots is useful taxonomically. The grouping based on the chromatograms agreed well with the Gibson grouping, but indicated that Cowans group III contained some dissimilar organisms.

Whole-cell hydrolysates were prepared from 58 strains of nocardiae and streptomycetes. Strains morphologically intermediate between the two genera and morphological variants of the same strains were included. Paper chromatograms made from the whole-cell hydrolysates clearly demonstrated meso -diaminopimelic acid as a major constituent of cultures of Nocardia spp., and LL-diaminopimelic acid as a major constituent of cultures of Streptomyces spp. In cultures of ten strains of N. madurae and of three of N. pelletieri, meso -diaminopimelic acid predominated, thereby supporting the assignment of these species to the genus Nocardia .

When purified diphtheria toxoid was subjected to filter paper electrophoresis a separation was achieved under certain conditions. The flocculating activity of the toxoid would appear to be localized in only one of the four fractions obtained. This fraction represented about one-third of the material used (0.05 ml.), thus indicating the relative impurity of a highly purified material.

Summary It was found that Vibrio comma phages absorbed on filter paper maintain their lytic property for a long period of time. A mixture of four phages covering a wide range of true cholera vibrios was prepared and absorbed on filter paper disks which were stored for 4 months. These disks proved helpful in the identification of V. comma.

A laboratory technique for measuring the mobility of solution on filter paper has been presented. Various exudates exhibited different mobilities on filter paper; this also applied to exudates of the Same origin (Pleura punctates.) When diluted, the exudates exhibited increased mobility, indicating that the concentrations of various exudate components were of importance. The addition of cells (5 × 106 and 25 × 106 P 388 D cells/ml exudate) did not significantly change the mobility, whereas the mobility of protein fractions (human albumin and gamma globulin) was inversely related to the concentration of the solutons. These finding may be of significance to the reliability of present methods used for the quantitation of gingival exudation.

The test paper here described makes it relatively easy to choose the antibiotic best suited for treatment of a given bacterial infection. Each type of test paper bears both a culture medium and an antibiotic and is supplied, dry and sterile, in a transparent envelope. For a given test, the paper is wetted and inoculated and the envelope with its contents is incubated. It is not necessary to wait for the appearance of visible colonies because the metabolic activity of the bacteria is detected by an indicator (triphenyltetrazolium chloride) which is colorless until reduced to a bright red dye by bacterial enzymes. The reliability of the new test was established by a comparative study with other methods in 100 clinical cases. The test paper is easily stored, and the entire unit can be incinerated after use. It is therefore useful in general office practice and should help to make sure that each patient receives the most effective antibiotic.

Paper strips impregnated with stains and chemicals for bacterial staining (Gugol Scientific Corporation, distributed by Wampole Laboratories) were evaluated. The quality of staining 100 Gram-positive and 80 Gram-negative microorganisms was compared with Hucker's modification. Over-decolorization of strip-stained Gram-positive organisms was the most frequent cause of inconsistency. Paper strip “acid-fast” stain was compared with the Ziehl-Neelsen technic for 90 mycobacteria and nine Nocardia strains. Mycobacteria and Nocardia were consistently well stained by the paper strip technic; however, Nocardia were not stained by the true acid-fast method. Ninety-five clinical smears, stripstained by physicians on the wards, were evaluated. Ease of performance and stability of reagents make the paper strip technics practical for use in clinics, wards, and small bncteriologic laboratories.

Ikawa, Miyoshi (University of New Hampshire, Durham), James W. Morrow, and Sheila J. Harney . Paper chromatographic system for the identification of glycerol in bacterial cell walls. J. Bacteriol. 92: 812–814. 1966.—The solvent system consisting of isopropanol-5% boric acid (7:1, v/v) separates glycerol from the other carbohydrate constituents which are found in hydrolysates of bacterial cell walls. This system is useful for the identification of glycerol even when anhydroribitol and rhamnose are both present, and has been found to be applicable on cell wall hydrolysates as well as on synthetic mixtures.

Bartholomew , J. W. (University of Southern California, Los Angeles), Thomas Cromwell, and Richard Gan . Analysis of the mechanism of Gram differentiation by use of a filter-paper chromatographic technique. J. Bacteriol. 90: 766–777. 1965.—Data are presented which demonstrate that the mechanism of gram-positivity could not be due solely to factors such as a single, specific gram-positive substrate, specific affinities of crystal violet for certain cellular components, a specific crystal violet-iodine-substrate complex, or to any specific characteristic of the dye, iodine, or solvent molecules. Ruptured cells of gram-positive organisms stain gram-negatively when subjected to a standard Gram-stain procedure. However, when stained fragments of broken cells were deposited in thick layers on the surface of filter-paper strips and exposed to decolorizers, the rate of dye release correlated with the Gram characteristic of the intact cell. Therefore, the intact cell in itself is not an absolute requirement for Gram differentiation. The data are interpreted as indicating that the mechanism of Gram differentiation primarily involves the rate of permeation of molecules (dye, iodine, solvent) through the interstitial spaces of cell-wall material.