bluuebunny/crossref_metadata_2025 huggingface dataset:
Electrophoretic Variants of Phosphoglucomutase in
Saccharomyces
Species
Authors: Douglas H. C. , Grindall D. Paul , Talbott Helen
Published: January 2020

Strains of
Saccharomyces cerevisiae
and species with which
S. cerevisiae
is interfertile display a characteristic pattern of electrophoretic variants of phosphoglucomutase (PGM) consisting of a major component and one or two minor components, all of which migrate toward the cathode. The patterns are consistent with an earlier finding that two unlinked genes, one of which has two known alleles, determine the synthesis of PGM in
S. cerevisiae
. The PGM patterns of strains of
S. fragilis, S. lactis
, and
S. marxianus
, species thought to be closely related to each other and only distantly related to
S. cerevisiae
, also displayed a characteristic pattern of PGM variants, but it was quite different from that of
S. cerevisiae
. In these species five or six electrophoretic variants could be detected, all of which migrated toward the anode. We interpret the differences in the PGM variants of the two groups of species as a reflection of differences in genetic composition which have arisen in two phylogenetically distinct groups that have become sexually isolated from each other.
Morphology and Round Body Formation in
Vibrio marinus
Authors: Felter R. A. , Colwell R. R. , Chapman G. B.
Published: January 2020

The morphology of
Vibrio marinus
MP-1 was studied by phase and electron microscopy. The ultrastructure of the vibrio form of
V. marinus
was found to be typically gram-negative with a trilaminar plasma membrane and cell wall. The coccoid or round bodies noted in otherwise pure cultures of
V. marinus
were frequently found in early and late stationary phase of growth. The round bodies in ultrathin section were found to contain at least one, and often three or four, cell units. Three types of round bodies were observed in ultrathin section, each differing in size and behavior: “spherules,” “spheres” or the “round body,” and “giant cells” or “macrospheres.” The round bodies appeared to be associated with, or to result from, the constrictive cell division of
V. marinus
.
Amino Acid Antagonist Death in
Escherichia coli
Authors: Rabinovitz M. , Finkleman A. , Reagan R. L. , Breitman T. R.
Published: January 2020

Six analogues of amino acids killed corresponding auxotrophs of
Escherichia coli
. With all but one analogue, protein synthesis was required for lethality.
Alternate Requirement for Vitamin B
12
or Methionine in Mutants of
Pseudomonas denitrificans
, a Vitamin B
12
-producing Bacterium
Authors: Lago Barbara D. , Demain Arnold L.
Published: January 2020

Experiments are described which indicate that
Pseudomonas denitrificans
, an organism that overproduces vitamin B
12
, uses the B
12
pathway exclusively for methionine synthesis.
Agar Plaque Formation by Mouse Spleen Cells in Response to Vaccination with Vi Antigen and Typhoid Vaccines
Authors: Esposito V. M.
Published: January 2020

Vi-containingSalmonella typhosaorganisms were resistant to killing by immune spleen cells in the presence of complement even when the cells were producing Vi antibody.
Effect of Oxygen on Heme and Porphyrin Accumulation from δ-Aminolevulinic Acid by Suspensions of Anaerobically Grown
Staphylococcus epidermidis
Authors: Jacobs N. J. , Jacobs J. M. , Sheng G. S.
Published: January 2020

The effect of various conditions on the accumulation of porphyrins and heme by resting suspensions of anaerobically grown cells of
Staphylococcus epidermidis
was examined. Anaerobically grown cells contain 10 to 15% of the amount of protoheme found in cells grown aerobically. Resting suspensions of anaerobically grown cells, when incubated aerobically in buffer with δ-aminolevulinic acid and glucose for 60 min, exhibited a fourfold increase in protoheme content. At high levels of δ-aminolevulinic acid, there was also a significant accumulation of porphyrins with the solubility and chromatographic properties of coproporphyrin and uroporphyrin. Protoporphyrin was not accumulated. When oxygen was excluded from the incubation mixture, accumulation of protoheme was prevented, but accumulation of coproporphyrin and total porphyrin was enhanced. Nitrate served as an electon acceptor as indicated by its reduction to nitrite; however, nitrate did not substitute for oxygen in causing the accumulation of protoheme. These results suggested that oxygen is required for one of the late steps of heme synthesis in
S. epidermidis
, possibly for the conversion of coproporphyrinogen to protoporphyrin. The inability of nitrate to substitute for oxygen suggests a role for molecular oxygen as a substrate rather than as an electron acceptor for heme synthesis.
Locus-specific Changes in Cell Wall Composition Characteristic of Osmotic Mutants of
Neurospora crassa
Authors: Livingston Laura R.
Published: January 2020

The osmotic phenotype of
Neurospora crassa
is characterized by inhibition of growth at high osmolalities of growth medium. Mutations at six osmotic loci of linkage group I were examined to assess the biochemical and physiological effects of these mutants. Isolated cell walls from 23 osmotic strains were compared with the wild type with regard to quantitative levels of the following components: percentage of total dry weight, total glucose, alkali-soluble glucose, nonglucose carbohydrates, amino acids, glucosamine, galactosamine, and a compound tentatively identified as quinovosamine. The last component has not previously been observed in
N. crassa
cell walls. Although the cell wall dry weight content of osmotic mutants was not altered, walls isolated from all of the osmotic strains had less alkali-insoluble glucose than those from the wild type. In addition, all of the loci except
cut
exhibited other consistent differences from the wild type. The
os-1, os-3
, and
os-5
mutants had low levels of alkali-soluble glucose. The
os-3
and
os-5
mutants had high levels of nonglucose carbohydrates, and
flm-2
had a low level of nonglucose carbohydrates. The
os-4
mutants had low levels of galactosamine and amino acids and high levels alkali-soluble glucose. An
os-1
mutant, B135, produced less of the whole alkali-soluble fraction of the cell wall.
Intracellular Protein Breakdown in a Thermophile
Authors: Epstein Israel , Grossowicz Nathan
Published: January 2020

Protein breakdown of 5 to 7% per hr was found in nitrogen-starved cells of an unclassified prototrophic thermophilic bacillus; a similar protein-breakdown rate (6.5% per hr) was found in resting cells of
Escherichia coli
. In the thermophile, the rate of protein breakdown was markedly influenced by the temperature; it was maximal between 45 and 55 C, and it decreased considerably at 35 and 75 C, temperatures which are only slightly below or above the minimal and maximal growth temperatures. Growing cultures of the thermophile showed little, if any, protein breakdown, a finding similar to that of others with
E. coli
.
Biosynthesis of Glucosyl Diglycerides by
Mycoplasma laidlawii
Strain B
Authors: Smith Paul F.
Published: January 2020

Monoglucosyl diglyceride is synthesized from 1,2-diglyceride and uridine-5′-diphosphoglucose (UDP); diglucosyl diglyceride from monoglucosyl diglyceride, and uridine-5′-diphosphoglucose by membranes of
Mycoplasma laidlawii
strain B. All of these enzymatic activities reside in the membrane. Membranes solubilized by detergent action or succinylation and acetone powders of membranes were inactive. Requirements for Mg
2+
, UDP, and appropriate lipid acceptor were demonstrated for biosynthesis of both glycolipids. Glucose-1-phosphate plus uridine triphosphate could replace the UDP requirement. A medium of relatively high ionic strength and a critical concentration of sodium lauryl sulfate stimulated biosynthesis of the monoglucosyl diglyceride. The optimal
p
H for both reactions was 8.0. A specificity for 1,2-diglyceride from the homologous organism was found for optimal synthesis of the monoglucosyl diglyceride, and a specificity for monoglucosyl diglyceride was found in the case of diglucosyl diglyceride synthesis. Both reactions were specific for UDP.
Effect of Bacterial Contamination on Cecal Size and Cecal Contents of Gnotobiotic Rodents
Authors: Loesche Walter J.
Published: January 2020

In the present investigation the effect of various bacterial contaminations of gnotobiotic mice and rats on cecal size is presented. Of the species tested,
Bacteroides oralis
and
Fusobacterium nucleatum
did not establish in germ-free mice.
Streptococcus mutans, Clostridium difficile
, a
Neisseria
strain and two recent cecal isolates established, but failed to exert an effect upon the cecum of mice. A group K streptococcus and
B. fragilis
increased the cecal size apparently by increasing the levels of water-soluble protein, peptides, and carbohydrates in the cecal contents. Mixed ileal bacteria decreased the cecal size by preventing accumulation of soluble proteins and carbohydrates in the cecum. A
Peptococcus
strain caused a reduction by lowering the levels of insoluble material in the cecum. When this strain was combined with two
Clostridium
isolates and introduced into gnotobiotic rats, 50 to 65% cecal reduction was observed. This polycontamination did not decrease the per cent water of the cecal contents but caused lower levels of both soluble and insoluble material to accumulate in the cecum. No net nitrogen absorption from the distal small intestine occurred in either the germ-free or polycontaminated rats.
Biosynthesis of Desthiobiotin in Cell-free Extracts of
Escherichia coli
Authors: Pai C. H.
Published: January 2020

Cell-free extracts of
Escherichia coli
were active in catalyzing the synthesis of a biotin vitamer from 7,8-diaminopelargonic acid. The vitamer was identified as desthiobiotin on the basis of its chromatographic and electrophoretic characteristics and its biotin activities for a variety of microorganisms. The reaction was stimulated five-fold by bicarbonate, suggesting that an “active CO
2
” was incorporated into the carbonyl carbon of desthiobiotin. The enzyme was demonstrable in a wild-type (K-12) and in all biotin mutants of
E. coli
that were tested, with the exception of a strain which was able to grow on desthiobiotin but not on diaminopelargonic acid. Furthermore, the enzyme was repressible by biotin in all of the strains tested. These results are consistent with the hypothesis that the biosynthesis of desthiobiotin from 7,8-diaminopelargonic acid is an obligatory step in the biosynthetic pathway of biotin in
E. coli
.
Growth and Sporulation of
Bacillus subtilis
Mutants Blocked in the Pyruvate Dehydrogenase Complex
Authors: Freese Ernst , Fortnagel Uta
Published: January 2020

Two “ACE” mutants of
Bacillus subtilis
which require acetate for growth on glucose minimal medium have been isolated. They do not grow with acetoin, 2,3-butanediol, fatty acids, isoleucine, lipoic acid, malic acid, pyruvic acid, succinic acid, thiamine, or valine, but respond somewhat to glutamate or citrate. The mutants lack the activity of the pyruvate dehydrogenase complex; they excrete pyruvate and later acetoin. They grow in nutrient sporulation medium (NSMP) to one-half the normal turbidity and do not sporulate subsequently. When acetate is added to NSMP (at the optimal concentration of 0.07
m
), the ACE mutants grow to the normal turbidity and then sporulate normally. Growth but not sporulation is restored in NSMP upon addition of 2,3-butanediol, citrate, glucose, glutamate, glycerol, or ribose, but not upon addition of acetoin, malate, oxaloacetate, pyruvate, and several other compounds. After growth in NSMP has stopped, the mutants incorporate uracil only at a very low rate, which can be increased by the addition of acetate, citrate, or glutamate. Furthermore, the metabolism of acetoin is prevented after growth has stopped but can be restored by the addition of acetate. All these results can be explained by a lack of reduced nicotinamide adenine dinucleotide (NADH) resulting from the deficiency in acetylcoenzyme A. In fact, after growth of the ACE mutants had stopped, the NADH concentration was at the borderline of measurability, whereas it increased significantly upon addition of glucose. The growing standard strain contains, at the same bacterial turbidity, at least 20 times more NADH (230 pmole/optical density unit at 600 nm) than the nongrowing ACE mutants. The isolated spores, obtained after growth in NSMP plus acetate, can be initiated to germinate in the presence of either
l
-alanine or the combination of
l
-asparagine, fructose, glucose, and potassium; addition of acetate is not required and has no effect.
Effects of Ultraviolet Radiation on Respiration and Growth in Radiation-resistant and Radiation-sensitive Strains of
Escherichia coli
B
Authors: Hamkalo Barbara A. , Swenson P. A.
Published: January 2020

Ultraviolet (UV) irradiation at 254 nm causes different respiration and growth responses in log-phase cultures of
Escherichia coli
B/r and B
s−1
. These differences are correlated with the ability and inability, respectively, of these bacterial strains to repair UV-induced lesions in deoxyribonucleic acid (DNA). After irradiation, B
s−1
cells (radiation-sensitive) exhibit uncoupling of growth and respiration; growth and synthesis cease, whereas respiration continues. B/r cells (radiation-resistant) grown on glycerol exhibit severe temporary inhibition of growth and respiration after UV, and the coupling of these two processes is maintained, except at a very high UV dose. Inhibition begins at about the time DNA synthesis resumes and continues for a period of time that is dependent upon dose. Glucose-grown cells do not exhibit severe respiratory, growth, and synthetic inhibitions; these processes remain coupled in the cells during the postirradiation period. Photoreactivation treatment delays uncoupling of growth and respiration in B
s−1
and prevents inhibition of respiration and growth in B/r. These results indicate that the postirradiation responses result from the presence of pyrimidine dimers in DNA. Ultraviolet irradiation of B/r and B
s−1
cells results in an accumulation of adenosine triphosphate by 30 min after UV. This accumulation decreases with time and does not appear to be related to the inhibition of respiration in glycerol-grown B/r cells. The results on B/r are interpreted in terms of a control mechanism for reestablishment of a balance among macromolecules in the irradiated cells so as to provide them with the potential to survive. The specific steps in such a reestablishment of balance appear to depend upon the substrate oxidized. In B
s−1
cells, which cannot repair UV-induced damage in DNA, some control mechanism that coordinates cellular processes may be inactivated.
Fine Structure of Fruiting Bodies of
Stigmatella aurantiaca
(
Myxobacterales
)
Authors: Voelz Herbert , Reichenbach Hans
Published: January 2020

The fruiting body of
Stigmatella aurantiaca
consists of a thick stalk supporting a number of individual cysts. The stalk is made of discontinuous tubules, of dimensions known for vegetative cells, which are oriented parallel to the longitudinal axis of the stalk. The red-brown cysts contain numerous, randomly oriented myxospores which are surrounded by thick, fibrous capsules. Their cell walls are wavy or ruffled and exhibit fewer budlike infoldings than reported for myxospores induced in liquid. We suggest that the extended time period available for metabolic and regulatory adjustments by the cell during morphogenesis within cysts accounts for the presence of considerably fewer deep cell wall infoldings than in glycerol-induced myxospores.
Cell Wall Residues in Yeast Protoplast Preparations
Authors: Bacon J. S. D. , Jones D. , Ottolenghi P.
Published: January 2020

Protoplast preparations made from
Saccharomyces cerevisiae
by prolonged treatment with snail digestive juice contained fibrils and chitinous bud-scar residues from the original cell wall.
Adaptation of Meningococcal L-Forms to Agar-free Liquid Medium
Authors: Stewart Ronald H. , Wright D. N.
Published: January 2020

Neisseria meningitidis
L-forms were adapted to an agar-free liquid medium after one passage in 0.05% agar-broth followed by two passes in 0.01% agar-broth.
Plaque Assay System for Several Species of
Rickettsia
Authors: McDade Joseph E. , Stakebake Jack R. , Gerone Peter J.
Published: January 2020

The plaque assay developed in this laboratory for the Bitter Root strain of
Rickettsia rickettsii
has recently been shown to be appropriate for other rickettsiae.
Comparison of three procedures for biochemical testing of anaerobic bacteria
Authors: Moore H B , Sutter V L , Finegold S M
Published: January 2020

The Analytab Products, Inc. (API), anaerobic multitest microsystem (MICRO) was compared with the Center for Disease Control conventional (CONV) thioglycolate (supplemented with hemin and vitamin K1) system and with pre-reduced anaerobically sterilized (PRAS) media as recommended by the Virginia Polytechnic Institute. Growth from a solid medium was suspended to produce standard inocula. Substrates included 16 carbohydrates, indole, urea, gelatin, and esculin. API strips were inoculated in air and incubated in GasPak (BBL) jars. MICRO tests were read at 1 and 2 days. CONV tests at 1, 2, and 7 days, and PRAS tests at 3 weeks. One hundred thirty well-characterized strains of anaerobes (76 gram-negative rods, 16 cocci, 26 gram-positive nonsporeforming rods, and 12 clostridia), including 48 reference strains, were studied. Of 2,600 tests performed, 2,085 (80.2%) showed agreement with all three methods. There was 90.9% agreement between the MICRO and CONV, 84.9% between the MICRO and PRAS, and 84.6% between the CONV and PRAS tests. All MICRO tests were reliable except for indole, which was not sensitive enough, and gelatin, which was very insensitive. The MICRO system permits performance of biochemical tests at the workbench in the average clinical laboratory without the need for expensive equipment and time-consuming procedures.
Reference equine antisera to 33 human adenovirus types: homologous and heterologous titers
Authors: Hierholzer J C , Gamble W C , Dowdle W R
Published: January 2020

Equine antisera to human adenovirus types 1 to 33 were prepared and evaluated by hemagglutination-inhibition and serum neutralization tests. Detailed data on the potency and purity of the immunizing antigens were tabulated as one means of evaluating the antisera. Most of the 52 hemagglutination-inhibition and 25 serum neutralization major or minor heterotypic responses among the equine antisera were observed at similar levels in previous studies with rabbit antisera and appeared to represent genuine antigenic relationships among the human adenoviruses. Equine antisera to human adenoviruses 1 to 33 and a similarly packaged normal horse serum served as lots of fully tested sera for definitive typing of isolates and as reference standards for evaluating other antisera.
New satellitism test for isolation and identification of Haemophilus influenzae and Haemophilus parainfluenzae in sputum
Authors: Evans N M , Bell S M , Smith D D
Published: January 2020

A new satellitism test designed to facilitate the isolation and identification of Haemophilus influenzae and Haemophilus parainfluenzae is described. In the basal medium, nicotinamide adenine dinucleotide is incorporated at a concentration of 0.2 mug per ml, an amount adequate for H. influenzae but not for H. parainfluenzae. Two disks are placed on the surface of the medium, one disk being impregnated with 60 mug of hemin and the other with 15 mug of nicotinamide adenine dinucleotide. Under these conditions, H. influenzae strains grow around the hemin disk only and the majority of H. parainfluenzae grow around the nicotinamide adenine dinucleotide disk. This procedure gives results which are more clear cut than other established methods, especially in sputum culture.
Biotypes of Proteus rettgeri
Authors: Penner J L , Hinton N A , Hennessy J
Published: January 2020

Examination of 729 isolates of Proteus rettgeri showed that 674 reacted positively in tests for phenylalanine deamination, indole production, growth on citrate, and acid production from meso-inositol, and negatively for L-ornithine decarboxylation and acid production from lactose, maltose, D-xylose, and L-arabinose. Only 51 isolates differed in one, and four differed in two of these ten reactions, which were taken as the core characteristics of the species. On the basis of additional tests (acid production from salicin, L-rhamnose, D-mannitol, adonitol, and D-arabitol), the 729 isolates could be separated into five groups. Goups 1, 2, 3, and 4 could be further separated on the basis of the reaction with meso-erythritol, and group 5 could be subdivided on the basis of reaction with D-mannitol. Two metabolically distinct kinds of P. rettgeri were recognized. Isolates of the first kind (groups 1, 2, 3, and 4) each utilized both adonitol and D-arabitol and most utilized meso-erythritol. Isolates of the other kind (group 5) were negative with the three polyhydric alcohols but resembled, in their reactions, some strains of Providencia stuartii. These may be intermediates between P. rettgeri that catabolize these substrates and the Providencia.
Oxygen tolerance of fresh clinical anaerobic bacteria
Authors: Tally F P , Stewart P R , Sutter V L , Rosenblatt J E
Published: January 2020

The oxygen tolerance and sensitivity of 57 freshly isolated anaerobic bacteria from clinical specimens was studied. All the organisms tolerated 8 h or more of exposure to oxygen in room air. Growth of the isolates in increasing oxygen concentrations demonstrated that the 57 isolates varied in oxygen sensitivity from strict to aerotolerant anaerobes. Comparison of the oxygen tolerance and sensitivity showed that the most tolerant organisms (best survival after prolonged exposure) included anaerobes capable of growth at only 0.4% or less O2 (strict) as well as those able to grow in as much as 10% O2. The least tolerant were predominately strict anaerobes. Decrease in the inoculum size from a concentration of 10(8) to 10(6) colony-forming units per ml had only a minor effect. The data indicate that the brief oxygen exposure with bench techniques in clinical laboratories would not be deleterious to the anaerobic bacteria present in clinical specimens.
Pagano-Levin Candida test medium: evaluation using vaginal samples
Authors: Sinski J T , Kelley L M , Reed G L
Published: January 2020

Since 1958 pagano-Levin medium (PLM) has been used as an aid in the identification of Candida albicans. However, no statistical analysis of its effectiveness based on large number of random human specimens has been reported. The present study compared PLM (now called Candida test) with Sabouraud's plus antimicrobials and Littman's ox-gall agar without antimicrobials. Of 500 random vaginal samples 24.8 were true positives, 71.9% were true negatives, 2.2% were false positives, and 1.2% were false negatives on PLM. If only samples identified as C. albicans were considered, 95.4% were true positives and 4.6% were false negatives. PLM did not inhibit C. albicans from the vagina. No one medium was found superior to the others for the purpose of isolating and identifying C. albicans. Of the five strains of C. parapsilosis, the only other Candida species isolated, all the samples grew on PLM but most were inhibited on Sabouraud's medium plus antimicrobials.
Improved recognition and quantitation technique for oncornaviruses
Authors: Stephens R , Traul K , Woolf D , Lowry G , Lelek J
Published: January 2020

A technique to recognize and quantitate oncornaviruses using perforated pointed BEEM capsules has been developed in our laboratory. Virus samples that presented problems in counting, or those which could not be evaluated at all by negative staining, could be clearly defined and counted using the thin-sectioning BEEM capsule technique. Perforation of the BEEM capsule allowed rapid infiltration of reagents into the tip of the virus pellet and made further manipulation and orientation unnecessary. The sensitivity of this technique, determined by making serial dilutions of viral concentrates, allows observation of as few as 5 times 10(5) virus particles per ml. Precision in counting by this technique varied only +/- 0.3 log in repeat aliquots of identical concentrated virus samples quantitated, making this a highly useful and reliable system.
Identification of streptococci: use of lysozyme and Streptomyces albus filtrate in the preparation of extracts for Lancefield grouping
Authors: Watson B K , Moellering R C , Kunz L J
Published: January 2020

A combination of lysozyme and Streptomyces albus filtrate has been shown effective in extracting group-specific antigen for all commonly occurring serologically groupable streptococci. A prospective comparison of this method with that of Rantz and Randall (1955) for grouping 761 clinical isolates has confirmed its accuracy, which in our hands exceeded that of the latter more complicated method of serogrouping. Its rapidity and simplicity and the relatively low cost of the reagents involved make it practical for routine use in clinical bacteriology laboratories.
Carbohydrate fermentation plate medium for confirmation of Neisseria species
Authors: Faur Y C , Weisburd M H , Wilson M E
Published: January 2020

A carbohydrate fermentation plate medium is described for rapid and reliable confirmation of Neisseria gonorrhoeae, N. meningitidis, and other Neisseria species. The medium is based on a modification of NYC (U.S. Patent 3,846,241) medium, originally designed for the isolation of pathogenic Neisseria (3). A total of 715 clinical isolates were tested for their carbohydrate fermentation reactions on the medium, in parallel with cystin-Trypticase agar medium. In 82% of the strains tested, results were in agreement on both media and 18% gave conflicting results. The NYC modification provides rapid and accurate fermentation patterns for use in routine confirmation procedures for Neisseria species.
Assessment of virus infectivity by the immunofluorescent and immunoperoxidase techniques
Authors: Hahon N , Simpson J , Eckert H L
Published: January 2020

The immunofluorescent and immunoperoxidase cell-counting techniques were comparable in precision and reproducibility for the quantitative assessment of influenza virus infectivity. The dose-response function was linear with each procedure, and comparable results were obtained for estimating neutralizing antibodies in antiviral serum.
Antibodies to Epstein-Barr virus in patients with cryptococcosis
Authors: Levine P H , Diamond R D , Reisher J I
Published: January 2020

Antibody levels to the Epstein-Barr virus, the etiological agent for heterophile-positive infectious mononucleosis, have been demonstrated in high titer in a number of lymphomas as well as infectious mononucleosis. Recent reports have suggested that the elevated antibody levels to Epstein-Barr virus may be the nonspecific result of disordered cell-mediated immunity. This study of patients with cryptococcosis was therefore undertaken to examine another disorder of known etiology associated with a defect in cell-mediated immunity. In this study we found that antibody levels in cryptococcosis patients, including a group specifically demonstrated to be anergic to a series of skin test antigens, were no different than those in matched normal controls. At the present time, therefore, it is unlikely that elevated antibody levels can be explained solely on the basis of depressed cellular immunity.
A More Complete Evaluation of the Color-Coded Antigen for the Automated Reagin Test
Authors: Lambert N. G. , Ste-Marie A. R.
Published: January 2020

A comparative study, on 500 samples, of the toluidine red antigen, one of the low-cost, color-coded antigens for the Automated Reagin Test, shows a greater sensitivity than the rapid plasma reagin card antigen.
Use of cetylpyridinium chloride and sodium chloride for the decontamination of sputum specimens that are transported to the laboratory for the isolation of Mycobacterium tuberculosis
Authors: Smithwick R W , Stratigos C B , David H L
Published: January 2020

A method is presented for the decontamination, liquefaction, and concentration of sputum specimens that are in transport more than 24 h. The method is inexpensive, and culture results compare well with those obtained with the accepted N-acetyl-L-cysteine and sodium hydroxide method for the isolation of tubercle bacilli. The working solution, 1% cetylpyridinium chloride and 2% sodium chloride, is mixed in equal volumes with sputum before the specimens are shipped. Tubercle bacilli remained viable after 8 days of exposure to this solution. Only Lowenstein-Jensen medium was used because the cetylpyridinium chloride in the inoculum remains active on 7H10 or other agar base media and partially inhibits the growth of tubercle bacilli.
Association of Pseudomonas cepacia with chronic granulomatous disease
Authors: Bottone E J , Douglas S D , Rausen A R , Keusch G T
Published: January 2020

Pseudomonas cepacia was recovered from a number of infected sites in three patients with chronic granulomatous disease of childhood. The organisms were identified on the basis of their oxidative utilization of a variety of carbohydrates and their positive beta-galactosidase and oxidase activities. They were resistant to most antimicrobial agents and moderately susceptible to chloramphenicol. Peripheral blood leukocytes isolated from two siblings with chronic granulomatous disease, including one of the patients in this series, failed to kill P. cepacia in vitro. Prolonged prophylactic and antimicrobial therapy may well have played a significant role in the colonization and infection of these patients with P. cepacia.
Photoreactivation of Pseudomonas cepacia after ultraviolet exposure: a potential source of contamination in ultraviolet-treated waters
Authors: Carson L A , Petersen N J
Published: January 2020

Cells of a naturally occurring strain of Pseudomonas cepacia grown in distilled water were exposed to ultraviolet radiation. Irradiated samples incubated on membrane filters or in fluid recovery media in the absence of light showed no evidence of dark repair mechanisms. When samples were exposed to fluorescent light ranging from 50 to 950 foot candles (538 to 10,222 lux) of illumination, apparent photo-induced repair of ultraviolet injury resulted in 1- to 4-log increases in viable cells recovered.
Micromethod for Serotyping Strains of
Mycobacterium avium
Authors: Thoen Charles O. , Jarnagin Jerald L. , Champion Margaret L.
Published: January 2020

A rapid and economical micromethod for serotyping strains of
Mycobacterium avium
is described. Eighty-four of 91 strains representing 20 serotypes were successfully identified.
Evaluation of tyrosine medium for the identification of Enterobacteriaceae
Authors: Sheth N K , Kurup V P
Published: January 2020

Use of the tyrosine medium in the identification of members of the family Enterobacteriaceae form the basis of this paper. Six hundred and twenty-three strains were tested for their property of tyrosine degradation. Three hundred and seventy-two strains were positive in 24 h and an additional eight strains were positive in 48 to 72 h. These positive strains consisted of all 344 strains of the Proteus-Providencia group, all 25 strains of Citrobacter diversus, and three strains of atypical C. freundii. Our findings suggest that tyrosine degradation is a useful property for separating out the Proteus-Providencia group and in specific identification of C. diversus.
Evaluation of the R/B and Minitek systems for identification of Enterobacteriaceae
Authors: Shayegani M , Hubbard M E , Hiscott T , McGlynn D
Published: January 2020

The R/B and Minitek systems for the identification of Enterobacteriaceae were evaluated, and the results were compared with those obtained by conventional methods. Both systems were rapid and allowed correct identification of about 85% of the 294 isolates (23 species) examined. The individual biochemical reactions showed an overall agreement of 92.6% and 93.1% for the R/B and Minitek systems, respectively.
Pigment production by Cryptococcus neoformans from para- and ortho-Diphenols: effect of the nitrogen source
Authors: Chaskes S , Tyndall R L
Published: January 2020

Cryptococcus neoformans produced pigments when p-diphenols were substrates in a glucose-amino acid-salts medium. The best substrates were 2.5-dihydroxybenzoic acid and 2,5-dihydroxybenzenesulfonic acid. In contrast to the cellular pigment production from o-diphenols (hydroxyl groups in the 2,3- or 3,4-position of phenyl ring), the p-diphenols (1,4- or 2,5-positions for the hydroxyl groups) produced large amounts of soluble pigments that diffused into the medium. When an optimal source of nitrogen (glutamine, glycine, and asparagine) was used, 89% of the C. neoformans strains produced pigments from p-diphenols. In contrast, 0 to 67% of the strains produced pigments when a suboptimal nitrogen source (proline, ammonium sulfate, ornithine, and methionine) was used. When glutamine-glycine-asparagine was the nitrogen source, 100% of the C. neoformans strains produced pigments from o0diphenols, whereas 77 to 100% of the strains produced pigment when proline-ammonium sulfate-ornithine-methionine was the nitrogen source. Cryptococcus species other than C. neoformans and all tested Candida species failed to produce pigments from any of the substrates except when hydroquinone was used. A combination of glutamine-glycine-asparagine and 3,4-dihydroxyphenylalanine allowed differentiation of colonies of C. neoformans from C. albicans in 3 to 6 days. These data showed that pigment production from o- and p-diphenols served as an excellent biochemical test for the identification of C. neoformans.
Rapid isolation of Legionella pneumophila from seeded donor blood
Authors: Dorn G L , Barnes W R
Published: January 2020

The laboratory isolation of Legionella pneumophila from seeded donor blood, using the lysis-centrifugation technique, is described. Time to pure culture isolate was 3 to 4 days.
Production of cells and viruses in a new multiple-tube tissue culture propagator
Authors: Corbeil M , Trudel M , Payment P
Published: January 2020

A novel method for large-scale culture of anchorage-dependent cells is described. Culture vessels were constructed by using roller bottles and Pyrex tubing. Parallel glass tubes, equally spaced by silicone rings and inserted into the roller bottles, provided an increased culture area. The roller bottles were fitted with a second opening for medium recirculation. Culture vessels of 1,000-, 4,000-, 8,000- and 16,000-cm2 growth area were constructed to propagate Vero, BS-C-1, primary chicken embryo, and IMH-P cells. These cultured cells were used to efficiently produce several viruses: rubella virus, measles virus, poliovirus, cytomegalovirus, and herpesvirus of turkey.
Enzyme immunoassay for detecting Brucella antibodies in cow's milk
Authors: Thoen C O , Pietz D E , Armbrust A L , Harrington R
Published: January 2020

An enzyme immunoassay (EIA) was developed for detecting Brucella antibodies in milk of cows infected with Brucella abortus. The enzyme immunoassay using heat-killed cells of B. abortus strain 19 was of comparable sensitivity to the Brucella ring test in detecting antibodies in milk of a reference positive control cow experimentally infected with B. abortus strain 2308 and in milk of 16 naturally infected cows from which B. abortus was isolated. No detectable enzyme immunoassay reactions were present in milk of 11 noninfected controls. The enzyme immunoassay is a procedure which can be readily automated so that screening tests for brucellosis could be conducted at a reference laboratory where uniform conditions can be maintained.
Clinical laboratory evaluation of automated microbial detection/identification system in analysis of clinical urine specimens
Authors: Isenberg H D , Gavan T L , Sonnenwirth A , Taylor W I , Washington J A
Published: January 2020

More than 4,000 clinical urine specimens were evaluated with an automated microbial detection/identification system compared to a standarized manual analysis and the routine modalities used in five peer-group laboratories. The comparison indicates that the automated system recognizes the nine groups of significant microorganisms in urinary tract infections in hospitalized patients with the same efficiency as a standarized manual method. The automated system's ability to enumerate the bacterial populations in the original clinical specimen attained a high degree of accuracy.
Biotyping of Escherichia coli by a simple multiple-inoculation agar plate technique
Authors: Buckwold F J , Ronald A R , Harding G K , Marrie T J , Fox L , Cates C
Published: January 2020

A nine-test system using multiple-inoculation agar plates for biotyping of Escherichia coli is described. Testing of 959 strains resulted in 78 biotypes. On repeated testing, 96% of 182 strains had identical biotypes or differed by only one test. This system provides satisfactory differentiation among strains and is reproducible. Precise standardization of inoculum size is not required. Multiple inoculation allows time and cost-efficient testing of large numbers of strains.
Phage heterogeneity of coagulase-negative staphylococci isolated in the United States and Sweden from bovine milk
Authors: Holmberg O , Lyon R H , Price D , Farnsworth R J
Published: January 2020

A Swedish bovine and a Dutch human phage set for coagulase-negative staphylococci were used to phage type coagulase-negative staphylococci isolated from bovine milk from Minnesota dairy herds. A comparison was also made of the deoxyribonuclease activity of coagulase-negative staphylococci isolated from bovine milk in Sweden and Minnesota. Of 133 Minnesota isolates, only one could be typed with the Swedish set and one by the Dutch set, whereas of 218 Swedish strains, 49 could be typed with the Swedish set and 7 by the Dutch set. A larger number of coagulase-negative isolates from Sweden were deoxyribonuclease positive (35%) than were the similar isolates from Minnesota (12%). These findings substantiate the marked heterogeneity of coagulase-negative staphylococci isolated from bovine udders. Results presented point to the usefulness of establishing regional phage sets for epidemiological investigations of coagulase-negative staphylococci in cattle. It is anticipated that at a later stage the regional phage sets will be coordinated internationally.
Rapid serogroup identification of Neisseria meningitidis by using antiserum agar: Prevalence of serotypes in a disease-free military population
Authors: Craven D E , Frasch C E , Mocca L F , Rose F B , Gonzalez R
Published: January 2020

Nasopharyngeal cultures from 414 Marines were plated directly onto antiserum agar containing the antibiotics vancomycin, colistin, and nystatin for meningococcal isolation and serogroup identification. Meningococci were isolated from 267 Marines, giving a carrier prevalence of 64.5%. A total of 58% of the isolates could be placed into serogroups; of these 22.3% were group B, 4.7% were group C, 25.7% were group Y, 24.3% were group W135, and 23.0% were group 29E. No serogroup A organisms were recovered. Serotyping by agar gel double diffusion was performed on 148 strains. More than 70% of these strains were nontypable, and the disease-associated serotype 2 was present only in two group Y isolates. The same 148 isolates were also classified by major outer membrane protein patterns after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Polyacrylamide gel electrophoresis pattern IV was most common among all serogroups. The data demonstrate the effective use of antiserum agar for meningococcal surveillance and document the frequency of specific serotypes and polyacrylamide gel electrophoresis types among carrier isolates obtained from a nonrecruit military population.
Radioimmunoassay system using a serovar-specific lipopolysaccharide antigen of Leptospira
Authors: Kawaoka Y , Naiki M , Yanagawa R
Published: January 2020

The serovar-specific main (TM) antigen of Leptospira interrogans serovar kremastos strain Kyoto was labeled with sodium boro[3H]hydride after 1 h of oxidation with periodate. When 50 ng of the labeled compound was employed, 50% of the compound was bound to the antibodies contained in 7.5 x 10(-2) nl of anti-kremastos Kyoto serum. Under this condition, only 9 ng of the homologous TM antigen was required for 50% inhibition of serovar kremastos Kyoto 3H-TM antigen-anti-kremastos Kyoto serum binding, whereas 5,000 times as much TM antigen of serovar hebdomadis, which belongs to the same Hebdomadis serogroup as serovar kremastos, was required for the same inhibition. The TM antigens from serovars pomona, icterohaemorrhagiae, and copenhageni, which belong to different serogroups, showed no inhibition in an amount of up to 3 x 10(3), 2 x 10(5), and 2 x 10(5) ng, respectively. In the inhibition study using the serovar hebdomadis TM antigen, inhibition values slightly fluctuated due to the antigen's insolubility. When the TM antigens were solubilized with 0.1% sodium taurodeoxycholate, the fluctuation in inhibition values was minimized, and the cross-reactivity of the serovar hebdomadis TM antigen with anti-kremastos Kyoto serum was diminished, while the inhibitory activity of the serovar kremastos Kyoto TM antigen was enhanced.
Enzyme-linked fluorescence assay: Ultrasensitive solid-phase assay for detection of human rotavirus
Authors: Yolken R H , Stopa P J
Published: January 2020

Enzyme-linked immunosorbent assay (ELISA) has proven to be a useful assay system for the direct detection of infectious agents. However, when the usual color-producing substrates are employed, relatively large amounts of substrate must be hydrolyzed by the bound enzyme before detection can be achieved. We attempted to improve the sensitivity of ELISA by utilizing a substrate that yields a fluorescent product on enzyme action. The enzyme-linked fluorescence assay (ELFA) based on this principle was approximately 100 times more sensitive than the corresponding ELISA or radioimmunoassay for the detection of human rotavirus in a standard stool suspension. In addition, the ELFA for human rotavirus was capable of detecting antigen in six specimens that were negative by ELISA. Five of these specimens were obtained late in the course of confirmed rotavirus infections. ELFA provides a simple, reliable, ultrasensitive method for the rapid detection of viral antigen.
Pseudomonas putrefaciens as a cause of septicemia in humans
Authors: Schmidt U , Kapila R , Kaminski Z , Louria D
Published: January 2020

Septicemia due to Pseudomonas putrefaciens was found in an elderly man with chronic leg ulcers. This organism is rarely cultured from human material and has been reported to cause skin and ear infections in only a few patients. Its potential for invasiveness is documented in this case for the fourth time.
Bacteriophage typing of coagulase-negative staphylococci
Authors: Jefferson S J , Parisi J T
Published: January 2020

Cultures comprising the 10 species of coagulase-negative staphylococci proposed by Kloos and Schleifer (J. Clin. Microbiol. 1:82--88, 1975) were typed with bacteriophages isolated from Staphylococcus epidermidis. Although only 10.5% were typable, 50% of those identified as S. epidermidis were typed. Cultures from patients with middle ear infections were also classified by this system and phage typed.
Serological grouping of meningococci and encapsulated Haemophilus influenzae strains by latex agglutination
Authors: Leinonen M , Sivonen A
Published: January 2020

The latex agglutination method, utilizing antibody-coated latex particles, was adapted for serogrouping of Neisseria meningitidis and serotyping of encapsulated Haemophilus influenzae strains from agar plates. It was found to give more clear-cut results than conventional slide agglutination. A 100% agreement with the antiserum agar method was found for all strains isolated from blood or cerebrospinal fluid. Many meningococcal strains from nasopharyngeal carriers are autoagglutinable, but some of these gave a positive reaction with the group B latex reagent, although they were negative by the antiserum agar method. The latex agglutination method has several advantages over others: the lack of autoagglutination, easy performance, easy interpretation, and very low consumption of antisera.
Evaluation of the modified Micro-ID system for identification of Enterobacteriaceae
Authors: Buesching W J , Rhoden D L , Esaias A O , Smith P B , Washington J A
Published: January 2020

Micro-ID is a system designed to identify the Enterobacteriaceae by utilizing reagent-impregnated disks for 15 biochemical tests. Since its initial evaluations, the system has undergone modification in formulation and in its computer data base. In a dual-center evaluation, 306 isolates of Enterobacteriaceae were tested: 145 common and typical isolates at the Mayo Clinic and 161 unusual or atypical isolates at the Center for Disease Control. Each laboratory also exchanged 50 cultures to test the system's reproducibility. Micro-ID correctly identified 142 (98%) of the common clinical isolates and 123 (76%) of the unusual or atypical organisms. However, in this latter group, three species tested were not in the system's data base. When these organisms were deleted from the analysis, 138 of 146 (95%) of the unusual or atypical isolates were correctly identified. Analysis of the 100 isolates identified in duplicate revealed 93% reproducibility of genus and species identification and 62% reproducibility of octal code numbers. Of the 31 strains with the same identification but different code numbers, 74% differed in only one biochemical test.
Enzyme-linked immunosorbent assay for measurement of antibodies against pneumococcal polysaccharide antigens: comparison with radioimmunoassay
Authors: Callahan L T , Woodhour A F , Meeker J B , Hilleman M R
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) for measuring antibodies against each of 14 polysaccharides in contemporary pneumococcal vaccine is described, and the findings of tests of paired sera from vaccinated human subjects are compared with those obtained by radioimmunoassay. The findings were in very poor agreement, and this appears to be due to the lesser ability of the ELISA procedure to measure antibody of low avidity. The ELISA procedure described here is not considered to be a satisfactory substitute for radioimmunoassay for measuring antibody responses to pneumococcal vaccine.
Comparison of direct and standard antimicrobial disk susceptibility testing for bacteria isolated from blood
Authors: Mirrett S , Reller L B
Published: January 2020

To determine the reliability of early antimicrobial susceptibility testing, we compared the results of direct and standard single-disk diffusion methods for 581 positive blood cultures processed routinely by the clinical microbiology laboratory. The direct procedure differed from the standard one only in that the 0.5 McFarland inoculum was prepared from 1 ml of turbid broth rather than five isolated colonies from a subculture plate. A major discrepancy in results was defined as a change from susceptible to resistant or vice versa according to interpretive standards for zone diameters, whereas a minor discrepancy was defined as a shift to or from the intermediate category when paired direct and standard tests were compared. The overall agreement between the two methods was 94.6% of 2,308 comparisons. There were 119 minor (5.2%) and 6 major (0.3%) discrepancies. The major discrepancies were seen with three strains of Staphylococcus epidermidis and one strain each of S. aureus, Escherichia coli, and Enterobacter sp. Direct susceptibility testing of positive blood cultures that were pure by gram-stained smear provided reliable results 24 to 36 h earlier than conventional procedures; therefore, we recommended this procedure to guide early antimicrobial therapy in patients with bacterial sepsis.