bluuebunny/crossref_metadata_2025 huggingface dataset:
False-Positive Anti-
Toxoplasma
Fluorescent-Antibody Tests in Patients with Antinuclear Antibodies
Authors: Araujo Fausto G. , Barnett Eugene V. , Gentry Layne O. , Remington Jack S.
Published: December 2020

The indirect fluorescent-antibody (IFA) method for diagnosis of toxoplasmosis is widely used and is considered to be as specific as the Sabin-Feldman dye test. After observing a patient with systemic lupus erythematosus (SLE) who had a positive toxoplasma IFA test but a negative dye test, we studied sera with high titers of antinuclear antibodies from 16 SLE patients and from 2 with rheumatoid arthritis for
Toxoplasma
antibodies in the immunoglobulin G and M (IgG and IgM) IFA tests and the dye test. Results of these tests were compared with titers of antinuclear antibodies, precipitating antibodies to single-strand deoxyribonucleic acid (DNA), and binding antibodies by use of DNA labeled with
3
H-actinomycin D. Of 18 patients, 11 had IgG and 4 had IgM IFA
Toxoplasma
antibodies; only 2 had antibodies detectable in the dye test. The immunofluorescence patterns in the
Toxoplasma
IFA test were indistinguishable from those obtained in patients with toxoplasmosis without antinuclear antibodies. Absorption of SLE sera with DNA did not result in a decrease in
Toxoplasma
IFA titers. When SLE sera were absorbed with live
T. gondii
, a marked drop in IgG IFA titer was observed as well as a decrease in titers of antinuclear antibodies and
3
H-DNA binding. Treatment of
Toxoplasma
cells with deoxyribonuclease and ribonuclease did not decrease their fluorescence. These results suggest that
T. gondii
nuclear antigens can absorb antinuclear antibodies but do not have exposed substrates for deoxyribonuclease. Tests in which organisms containing “nuclear” antigens for IFA detection of antibodies to these organisms are used may result in “false-positives” with sera containing antinuclear antibodies.
Incorporation of Chlorine-36 into Ochratoxin A
Authors: Wei Ru-Dong , Strong F. M. , Smalley E. B.
Published: December 2020

Incubation of
Aspergillus ochraceus
NRRL 3174 in a medium containing Na
36
Cl incorporated
36
Cl into ochratoxin A. The highest incorporation (0.75 and 0.70%, respectively) was obtained when the radioactive chloride was added to the medium by the 2nd or 3rd day of incubation.
Characterization of an Unusual Strain of
Proteus rettgeri
Associated with an Outbreak of Nosocomial Urinary-Tract Infection
Authors: Traub W. H. , Craddock M. E. , Raymond E. A. , Fox M. , McCall C. E.
Published: December 2020

An outbreak of nosocomial urinary-tract infection was caused by a strain of
Proteus rettgeri
that fermented lactose overnight and was resistant to all antimicrobial drugs tested. The nonmotile isolates shared an O (somatic) antigen that differed from those of wild-type
P. rettgeri.
The organisms proved markedly serum-sensitive. In rats, the isolates elicited an acute interstitial nephritis with associated transient bacteriuria. Attempts to transfer the lac
+
trait and drug-resistance markers to recipient strains of
Escherichia coli
K-12 failed; exposure of the isolates to acridine orange yielded small numbers of non-lactose-fermenting variants which, however, were still as drug-resistant as before. Epidemiological studies failed to uncover the source of this unique strain and appeared to indicate exogenous spread of infection.
Scopafungin, a Crystalline Antibiotic Produced by
Streptomyces hygroscopicus
var.
enhygrus
var. nova
Authors: Johnson LeRoy E. , Dietz Alma
Published: December 2020

Scopafungin (U-29,479) is a crystalline, nonpolyenic antimicrobial agent obtained from the culture broth of
Streptomyces hygroscopicus
var.
enhygrus
var. nova UC-2397. Scopafungin inhibits, in vitro, a variety of pathogenic fungi, yeasts, and gram-positive bacteria.
Zonal-Centrifuged Purified Duck Embryo Cell Culture Rabies Vaccine for Human Vaccination
Authors: Lavender J. F. , Van Frank R. M.
Published: December 2020

Rabies virus produced in duck embryo cell culture was concentrated from volumes of 14 to 30 liters to 400 to 800 ml by zonal centrifugation. Virus titers of peak fractions were from 100- to 1,000-fold greater than those of the starting material. Vaccines were prepared by combining fractions with peak virus titers and diluting back to 10 times concentration. The resulting β-propiolactone-inactivated vaccines, when prepared as lyophilized vaccines with AlPO
4
adjuvant diluents, were low in protein nitrogen (0.01 mg/ml), and three of four lots passed the National Institutes of Health potency test when tested as equivalent to a standard 10% suspension of duck embryo or mouse brain tissue vaccine. These vaccines also induced good sero-conversion in adult rabbits after a single 1-ml dose of vaccine. Guinea pigs sensitized with zonal-centrifuged purified duck embryo vaccine (with AlPO
4
adjuvant) did not exhibit anaphylactic shock reactions when challenged with homologous vaccine. Also, no anaphylactic shock reactions were observed when guinea pigs were sensitized with either a 10% experimental duck embryo vaccine or cell culture vaccine and then challenged with the zonal-purified vaccine. However, guinea pigs sensitized with cell culture or zonal-purified vaccine and then challenged with the 10% experimental vaccine did show slight transitory congestion. The 10% experimental whole duck embryo vaccine was responsible for all observed anaphylactic shock reactions whether homologous or heterologous.
Biochemical Properties of Haploid and Diploid Strains of
Penicillium chrysogenum
Authors: Pathak S. G. , Elander R. P.
Published: December 2020

An intensive parasexual genetics program in which industrial strains of
Penicillium chrysogenum
were used culminated in the isolation of a number of heterozygous diploid strains. The diploid clones were selected from heterokaryons formed from matings between mutant strains having complementary biochemical and conidial color markers. Several diploid cultures were compared with their haploid wild-type parents and other distantly related production strains on the basis of a variety of cultural and physiological criteria. The diploid strains characteristically produced conidia of larger volume and higher deoxyribonucleic acid content. Some were vigorous with respect to growth rate and onset and degree of conidiation. One diploid strain (WC-9) had a 46% greater oxygen uptake rate and oxidized glucose at a 57% greater rate than its haploid parent (M-2). It also produced 33% higher concentrations of β-galactosidase, 66% more alkaline protease, and 53% more glucose oxidase than the M-2 haploid parent. The selection of rare stable diploid mold cultures through the use of parasexual genetics offers a unique approach to the direct selection of mutants with potential for increased enzyme formation.
Dose Responsiveness and Variation Among Inbred Strains of Mice in Production of Interferon After Treatment with Poly (I)· Poly (C) [Poly-
d
-Lysine] Complexes
Authors: Rice J. M. , Turner W. , Chirigos M. A. , Spahn G.
Published: December 2020

Poly-
d
-lysine forms a stochiometric complex with poly(I) · poly(C) which has a higher T
m
(83 C in 0.15
m
NaCl) than the uncomplexed double-stranded polyribo-nucleotide (63 C). The complex was superior to poly(I) · poly(C) alone as an interferon inducer in vivo. Significant serum interferon titers were attained in Swiss mice during a 24-hr period after intraperitoneal injection of 10 to 300 μg of poly(I) · poly(C) [1.0 poly-
d
-lysine] complex, at concentrations of 100 to 1,000 μg/ml. The serum interferon responses (average and maximum titers) of a series of inbred strains of mice to a single intraperitoneal injection of 100 μg of complex decreased in the order: Swiss > DBA/2 > C3H > BALB/c > CF-1 > AKR, C57Bl/6, NZB > SJL > NZW and varied by a factor of approximately 100 from the most to the least responsive.
High Aflatoxin Production on a Chemically Defined Medium
Authors: Reddy T. V. , Viswanathan L. , Venkitasubramanian T. A.
Published: December 2020

Aspergillus parasiticus
ATCC 15517 produced 28 to 30 mg of aflatoxin per 100 ml of a medium containing sucrose, asparagine, and salts in stationary and shaken cultures. In the absence of asparagine in the medium, the toxin yields fell drastically, and the thin-layer chromatograms of the chloroform extracts of the cultures indicated the total absence of aflatoxin G
1
and the presence of new intense blue and green fluorescent bands having

R
F

values lower than aflatoxins. Initial
p
H was critical and had to be around 4.5 for good growth and high toxin production on this medium. Optimum concentrations of KH
2
PO
4
and MgSO
4
·7H
2
O in the medium were much lower than those normally used in fungal growth media.
Repair of Injury in Freeze-Dried
Salmonella anatum
Authors: Ray B. , Jezeski J. J. , Busta F. F.
Published: December 2020

Repair of injury induced by freeze-drying
Salmonella anatum
in nonfat milk solids occurred rapidly after rehydration. Injury in surviving cells was defined as the inability to form colonies on a plating medium containing deoxycholate. Death was defined as inability to form colonies in the same medium without this selective agent. The rate of repair of injury was reduced by lowering the temperature from 35 C to 10 C and was extremely low at 1 C. Repair was independent of influence of
p
H between 6.0 and 7.0. Repair did not require synthesis of protein, ribonucleic acid, or cell wall mucopeptide, but did require energy in the form of adenosine triphosphate (ATP) synthesized through oxidative phosphorylation. The requirement for ATP was based on dinitrophenol or cyanide interference with repair. Dinitrophenol activity was
p
H-dependent; no repair occurred at
p
H 6.0 and some repair was observed at
p
H 6.5 and above. Injured cells were extremely sensitive to low concentrations of ethylenedinitrilotetraacetate. This indicated that freeze-drying injury of
S. anatum
may involve the lipopolysaccharide portion of the cell wall and that repair of this damage requires ATP synthesis.
Characterization of a Heat-Shock Process for Reduction of the Nucleic Acid Content of
Candida utilis
Authors: Ohta S. , Maul S. , Sinskey A. J. , Tannenbaum S. R.
Published: December 2020

A process for reducing the nucleic acid content of
Candida utilis
NRRL Y900 has been developed. The optimal process consists of heating the cells suspended in spent medium initially at
p
H 4.0 for various times at three different temperatures. Initially a heat-shock at 68 C for 1 to 3 sec is performed followed by incubation for 1 hr at 45 to 50 C and for a 2nd hr at 52 to 55 C. The distribution of degradation products has been characterized. Initially 90% of the nucleic acids were in a polymerized form (extractable by hot perchloric acid). After 30 min, much of this material was hydrolyzed but remained within the cell (extractable by cold perchloric acid). After 2 hr, most of the hydrolysis products leaked into the surrounding medium with only a small amount of low-molecular-weight material remaining within the membrane. Predominantly 3′-mononucleotides accumulated within the cell and eventually leaked from the cell.
Rapid Identification and Typing of Herpes Simplex Virus Types 1 and 2 by a Direct Immunofluorescence Technique
Authors: Nahmias A. , delBuono I. , Pipkin J. , Hutton R. , Wickliffe C.
Published: December 2020

An immunofluorescence (FA) technique has been developed which can identify herpes simplex virus (HSV) in clinical specimens and also type the virus directly as type 1 or type 2. This test, first applied to cervicovaginal specimens obtained from 80 mice genitally inoculated with HSV, indicated a sensitivity approaching 80% in comparison to standard viral isolation methods. A similar sensitivity was found when the test was applied to 185 clinical specimens with adequate cells for staining, which were obtained from a variety of sites of patients with suspect herpetic infection. In only 1 of 6 specimens positive by both FA and culture methods was the HSV type wrongly identified by the FA technique. There were also six specimens which were negative by culture methods but positive by the FA test, indicating a specificity of 91%. It is likely that these are not instances of false-positive tests but of other factors which may have resulted in negative viral isolations by culture methods. As more specific reagents become available, it is anticipated that the FA technique will have wider usage in diagnostic laboratories for the identification and typing of HSV types 1 and 2.
Simultaneous Use of Contrasting Fluorochromes to Separate Measles and Canine Distemper Viruses in a Common System
Authors: Gourlay James A. , Pemberton John R.
Published: December 2020

Measles and canine distemper viruses were grown together in a Vero monkey kidney cell line. Each virus could be identified and individually titrated by using the color contrast produced by the reddish tetra-methyl rhodamine isothiocyanate-tagged antimeasles conjugate and the green fluorescein isothiocyanate-tagged antidistemper conjugate. Both blue light and green light were used for the excitation of the fluorochromes. Incident light was transmitted to the specimen by a vertical illuminator of the Ploem type.
Rubella Virus Hemagglutination with a Wide Variety of Erythrocyte Species
Authors: Schmidt Nathalie J. , Dennis Juanita , Lennette Edwin H.
Published: December 2020

In HEPES diluents, rubella antigens agglutinated a wide range of species of erythrocytes, and the hemagglutination reaction showed little or no dependence on low temperatures.
Mechanical Method of Inoculating Plates for Antibiotic Sensitivity Testing
Authors: Kunz Lawrence J. , Moellering Robert C.
Published: December 2020

A mechanical method of inoculating culture plates for antibiotic sensitivity testing is described. This method, which involves the use of a modified laboratory centrifuge, is rapid and provides a homogeneous distribution of organisms.
Isolation of Inhibitory Factor in Raw Milk Whey Active Against Propionibacteria
Authors: Vedamuthu E. R. , Washam C. J. , Reinbold G. W.
Published: December 2020

Preparative isolation of the active component(s) in skim milk whey inhibitory for propionibacteria was made by using (NH
4
)
2
SO
4
salt fractionation. The crude preparation was further purified by Sephadex G-100 column separation. Disc-gel electrophoresis of the active peak from the Sephadex elution pattern (peak I) showed that this fraction contained almost all of the immune globulin in the column sample. The biologically inactive peaks did not contain any immune globulin. Starch-gel electrophoresis of the active peak revealed the presence of three separate immune globulin fractions. A correlation was also observed between hemolytic reaction of propionibacterial strains and relative resistance to whey inhibition. The investigation showed that one of the immune globulins of milk, pseudoglobulin, was mainly responsible for the suppressive activity of whey.
Comparison of Two Commercially Available Media for Detection of Bacteremia
Authors: Washington John A.
Published: December 2020

An analysis of 3,795 positive blood cultures obtained from 1,718 patients in a 2.5-year evaluation of Tryptic Soy Broth (TSB) and Thiol Broth is reported. Isolation rates of
Actinobacillus
and
Pseudomonas
were significantly greater in TSB, whereas isolation rates of
Streptococcus
and
Corynebacterium
(aerobic and anaerobic) were significantly greater in Thiol. Otherwise, the two media were similar. Disregarding contaminants, anaerobic bacteria represented 11% of positive cultures and 20% of patients with bacteremia. Eleven per cent of the patients had polymicrobial bacteremia.
Effect of Rapid Intravenous Infusion on Serum Concentrations of Amphotericin B
Authors: Fields Branch T. , Bates Joseph H. , Abernathy Robert S.
Published: December 2020

The magnitude of the concentrations of amphotericin B produced in serum of patients with systemic mycoses may significantly influence the outcome of therapy with this drug. Since amphotericin B is conventionally administered in intravenous infusions lasting 4 to 6 hr, we asked whether faster infusions of this drug might yield higher serum concentrations without an increase in dose. This question was studied in three patients who received 16 infusions of this drug: eight infusions administered slowly (5 hr) and eight administered rapidly (45 min). Serum concentrations after each rapid infusion were compared with those after a slow infusion administered to the same patient. The mean serum concentration of amphotericin B 1 hr after the rapid infusions (2.02 μg/ml) was significantly higher (
P
< 0.001) than the mean serum concentration of amphotericin B 1 hr after the slow infusions of this drug (1.18 μg/ml). Mean serum concentrations 18 and 42 hr after rapid infusion remained slightly but not significantly higher than respective mean concentrations after slow infusions. By yielding higher initial serum concentration, rapid intravenous infusion may be therapeutically more effective than slow infusion of amphotericin B. Although rapid infusions caused no more toxicity than did slow infusions, the lack of greater toxicity with rapid infusion of amphotericin B should be further documented prior to extensive clinical application of this procedure.
Microagglutination Procedures for Febrile Agglutination Tests
Authors: Gaultney Jerry B. , Wende Reuben D. , Williams Robert P.
Published: December 2020

Febrile agglutination tests were done by using as antigens
Brucella abortus, Salmonella
group D,
Proteus
OX19, and
Pasteurella tularensis
. Comparison of results from 23 sera showed that the microtechnique, rapid slide, and test tube methods gave similar titers, although those from the microtechnique were generally higher. The sensitivity of the microtechnique depended upon the concentration of antigen, and, to obtain reproducible results, the optimal concentration of antigens had to be determined by preliminary titrations against specific, positive control antisera. Readability of reactions in the microtechnique was enhanced by adding the dye Safranin O to diluent for antigen and by use of V-type, rather than U-type, microtiter plates. Tests were also done to determine the effects of dye and salt concentrations,
p
H, and temperature of incubation upon the titer of agglutinations by the microtechnique. Our results indicated that the microtechnique could be used for agglutination tests involving febrile antigens. The procedure is less time-consuming than the tube method and requires less antigen and serum than the latter method or the rapid slide method.
Dye Uptake Assay: an Efficient and Sensitive Method for Human Interferon Titration
Authors: Pidot Anne L. R.
Published: December 2020

Currently, human interferon (IF) assays are generally performed by plaque reduction or visual cytopathic effect methods. Both are time-consuming, subjective in interpretation, and, in the case of the latter, relatively insensitive. An adaption of the dye uptake method for human IF titration which uses foreskin-derived fibroblasts and vesicular stomatitis virus is described. This assay is reproducible and sensitive (1 unit = 3 international units). By direct comparison, however, it is somewhat less sensitive than the plaque reduction assay (1 unit = 1.1 international units). This assay is especially recommended for use in needed clinical investigations of human IF because of its technical simplicity, allowing efficient handling of large numbers of specimens.
Detection of R Factors in Naturally Occurring
Aeromonas salmonicida
Strains
Authors: Aoki Takashi , Egusa Syuzo , Kimura Takahisa , Watanabe Tsutomu
Published: December 2020

R factors were detected in
Aeromonas salmonicida
strains isolated from diseased salmonid fish. One of such R
+
strains was the one isolated in the United States as early as 1959.
Solaniol, a Toxic Metabolite of
Fusarium solani
Authors: Ishii Kenji , Sakai Kosei , Ueno Yoshio , Tsunoda Hiroshi , Enomoto Makoto
Published: December 2020

Fusarium solani
M-1-1 isolated from moldy bean hulls produces T-2 toxin, diacetoxyscirpenol, and a new toxic trichothecene, solaniol, in Czapek-Dox-peptone medium.
Simple, Reliable Cold Tray for the Recovery and Examination of Thermosensitive Organisms
Authors: Wiebe William J. , Hendricks Charles W.
Published: December 2020

A simple chamber is described for the isolation and handling of thermosensitive bacteria on board ship and in conventional research or classroom laboratories.
Fate of Carbon Passing Through the Glucose Pool of Rumen Digesta
Authors: Walker D. J. , Monk P. R.
Published: December 2020

The metabolism of the free glucose pool in rumen digesta from sheep fed roughage rations was studied by adding an insignificant quantity of glucose as uniformly labeled
14
C-glucose of high specific activity to in vitro incubation systems. In all experiments wherein only trace quantities of glucose were added to digesta, most of the
14
C-glucose entered acetate. This was true whether label was presented either as a single dose or by continuous addition over a period of 2 hr. Digesta collected at all times after feeding either once daily or at hourly intervals gave similar glucose dissimilation patterns. If, however, a relatively large quantity of carrier glucose was added together with the tracer, the
14
C-acetate:
14
C-propionate ratio was reduced by a factor of about 10. Physical removal of most of the protozoa from digesta generally had little effect on the dissimilation of
14
C-glucose added in tracer amounts, but in one experiment there was a decreased turnover of the free glucose pool and a marked reduction in
14
C entering butyrate. The paucity of
14
C entering propionate when only trace amounts of glucose were added to digesta suggests that this acid was largely formed from substrates whose carbon did not equilibrate with that in free glucose or with that in intermediates of free glucose metabolism.
Application of Enzyme Production Properties in Subtyping of Group A Streptococci According to T Type
Authors: Ofek I. , Fleiderman S. , Bergner-Rabinowitz S. , Ginsburg I.
Published: December 2020

The production of extracellular nicotinamide adenine dinucleotide glycohydrolase (NADG) and the cell-bound lipoproteinase (serum opacity reaction, SOR) by strains of different serological types of group A streptococci, in relation to the T typing, was studied. The production of both NADG and SOR, or only one of them, was found to be characteristic of serotypes, as determined by M and T antigen. No difference in the production of these enzymes was found in relation to M-positive and M-negative variants. Investigation into NADG and SOR production as related to the T type enabled the division of a single agglutination pattern into four main groups, each of which corresponds to one specific M type or more. Of the 370 strains belonging to 12 different T-agglutination patterns, 21% produced both enzymes and 42.5% failed to produce any of them, whereas the remaining 36.5% produced only one out of the two enzymes. Five streptococcal types which did not produce NADG and SOR also failed to synthesize streptolysin S at the early logarithmic phase of growth, indicating that streptolysin S production by young cultures may be also related to serotype. No correlation was found between the production of NADG-SOR as related to serotype and the production of streptolysin O, acid phosphotase, esterase,
N
-acetylglucosaminidase, hyaluronidase, streptokinase, and the cell-sensitizing factor. The practical and potential usefulness of NADG and SOR production in epidemiological studies is discussed.
Indirect Hemagglutination with
Mycoplasma
Antigens: Effects of
p
H on Antigen Sensitization of Tanned Fresh and Formalinized Sheep Erythrocytes
Authors: Krogsgaard-Jensen Anders
Published: December 2020

An investigation of the influence of different factors affecting the sensitivity of the indirect hemagglutination test has been performed with antigens of four mycoplasmas isolated from sheep or goat. Tanned erythrocytes of sheep, fresh and formalinized, were sensitized with the above antigens. It was demonstrated that, with formalinized erythrocytes, the sensitivity was increased by 50 to 100 times when the sensitization was done at a low
p
H level. The
p
H level was unimportant for sensitizing fresh erythrocytes. The greatest sensitivity of the indirect hemagglutination test was obtained with fresh rather than formalinized erythrocytes. Three different types of antigens were used, and the most suitable antigen was found to be the supernatant fluid from an ultrasonically treated centrifuged
Mycoplasma
suspension.
Estimation of the Growth of the T
1
Strain of
Mycoplasma mycoides
in Tryptose Broth by the Measurement of Lactate Dehydrogenase
Authors: Boarer C. D. H. , Read W. C. S. , Read J. Julie
Published: December 2020

A highly significant correlation coefficient (
r
= 0.97,
n
= 18) was found between the concentration of lactate dehydrogenase measurable after the organisms had been disrupted and the concentration of colony-forming units during the logarithmic phase of growth of a broth culture of the T
1
strain of
Mycoplasma mycoides
var.
mycoides.
A concentration of 4.60 × 10
-7
milliunits of lactate dehydrogenase for each colony-forming unit was established. This relationship was used to convert the concentration of lactate dehydrogenase in the culture into an estimate of the concentration of viable mycoplasma. The lactate dehydrogenase was estimated by following the oxidation of reduced nicotinamide adenine dinucleotide, in the presence of pyruvate substrate, at 366 nm in a spectrophotometer. The nicotinamide adenine dinucleotide oxidase system probably contributed a small amount of enzyme activity to the test when lactate dehydrogenase was measured in this way. The method has been described and evaluated for the estimation of titers from 10
7
to 5 × 10
9
colony-forming units per ml.
Rifampin-Blood-Agar as a Selective Medium for the Isolation of Certain Anaerobic Bacteria
Authors: Sutter Vera L. , Sugihara Paul T. , Finegold Sydney M.
Published: December 2020

A selective medium which allows detection of relatively small numbers of
Fusobacterium varium
in fecal specimens is described. Blood-agar containing 50 μg of rifampin per ml inhibits the growth of many species of
Bacteriodes
and of
F. fusi-forme/nucleatum
but allows good growth of
F. varium
and most strains of
F. mortiferum.
Quantitative cultures of 11 fecal specimens were done on rifampin and other selective and nonselective media.
F. varium
was recovered in counts of 10
6
and 10
7
per gram from two specimens on rifampin only. A third specimen yielded 10
10
F. varium
on several media, including rifampin. Some
Eubacterium
and
Clostridium
species also grew on rifampin, and these ordinarily were distinguished from the
Fusobacterium
by colony morphology. This medium is of value in fecal flora studies and should be useful with other kinds of specimens where mixtures of organisms are common.
Rapid Identification of Viruses by Indirect Immunofluorescence: Isolation and Identification of Adenovirus Types 4 and 7 and Coxsackievirus Type A21 in Microcultures
Authors: Stevens T. D. , Watkins H. M. S.
Published: December 2020

The indirect fluorescent-antibody test (IFAT), employing treated and standardized antiserum in a single pool, was earlier reported to have been used successfully for the preliminary identification of nine respiratory viruses in second to ninth passage multiplying in cells propagated on microscope slides. This report describes parameters affecting the isolation in first passage and the identification by IFAT of adenovirus types 4 and 7 and coxsackievirus type A21 present in stored clinical specimens inoculated into microcultures of WI-38 cells. Isolation frequency was comparable to that obtained in tube cultures, and identification by IFAT of viruses in microcultures could be accomplished in 3 to 4 hr after recognition of a cytopathogenic effect.
Bacteriological Evaluation of Two Test Methods for Chlorine in Swimming Pools
Authors: Savage Timothy B. , Stratton Lewis P.
Published: December 2020

It was found that orthotolidine and plastic test strips containing a mixture of syringaldazine and vanillinazine gave different results as to chlorine content of water samples containing urine or large amounts of bacteria while giving consistently similar results with water samples having only sodium hypochlorite added. Orthotolidine was found to give erroneously “safe” readings when in fact viable bacteria were observed on membrane filters after filtration.
Interferon Production and Host Resistance to Type II Avian (Marek's) Leukosis Virus (JM Strain)
Authors: Hong Chou C. , Sevoian Martin
Published: December 2020

Interferon production in both susceptible S- and resistant K-line chickens infected with type II leukosis virus of JM strain and turkey herpesvirus was studied. The resistant line of chickens produced higher levels of interferon than did the susceptible with JM virus infection during the experimental period. When both susceptible S-and resistant K-line chicks were vaccinated with turkey herpesvirus, the interferon production was quantitatively similar in the two lines.
Microbiological Evaluation of Protected Environments During Patient Occupancy
Authors: Bodey Gerald P. , Johnston Dianne
Published: December 2020

Microbiological monitoring has been conducted in two life island (LI) units and two laminar airflow (LAF) rooms while they were occupied by patients undergoing cancer chemotherapy. There were only 5 organisms per 1,000 ft
3
of air sampled in LAF rooms, 31 organisms in LI units, and over 3,000 organisms in regular hospital rooms. None of the floor samples obtained from hospital rooms was sterile, compared to over 70% in LAF rooms. The rate of deposition of organisms onto settling plates was one organism per 4.5 hr in LAF rooms compared to one organism per 0.08 hr in hospital rooms. Potential pathogens were isolated much more frequently from environmental samples obtained from hospital rooms than from LI units or LAF rooms. Two sites of persistent contamination arose in the LAF rooms: the vinyl tile flooring and the water supply system. Over half of the potential pathogens cultured from the protected environment units were cultured initially from the patients who occupied the units.
Interfering Effect of Incubation in Carbon Dioxide on the Identification of Pneumococci by Optochin Discs
Authors: Ragsdale A. Ruth , Sanford Jay P.
Published: December 2020

Incubation of pneumococci in a CO
2
environment decreases the zone sizes produced by optochin discs, with the result that pneumococcal-like organisms require further study.
Therapy of Staphylococcal Infections in Monkeys
Authors: Carlisle Harold N. , Saslaw Samuel
Published: December 2020

Intravenous inoculation of a penicillin-resistant phage type 80/81 staphylococcus caused lethal infection in seven of eight untreated monkeys. Daily intragastric administration of 50 mg/kg given in two equal morning and afternoon doses of cyclacillin and nafcillin was followed by mortalities of four of four and two of four monkeys, respectively. After 100 mg per kg per day, three of four and one of four monkeys receiving cyclacillin and nafcillin, respectively, died. Thus, mortality in controls and cyclacillin-treated monkeys was seven of eight as compared to three of eight after nafcillin treatment. Although the staphylococcus was more resistant to cyclacillin (minimal inhibitory concentration = 7.80 μg/ml) than to nafcillin (minimal inhibitory concentration = 0.31 μg/ml), regular rapid absorption and high levels of the former suggested potential efficacy. However, the similar mortality in cyclacillin-treated and control monkeys indicated that the in vitro data did not, in this instance, conform to the in vivo observations.
Propagation and Purification of High-Titer Human Cytomegalovirus
Authors: Chambers R. W. , Rose J. A. , Rabson A. S. , Bond H. E. , Hall W. T.
Published: December 2020

High-titered yields of human cytomegalovirus (CMV), strain AD 169, were produced in WI-38 cells in large roller bottles. Maximum plaque titers were observed by the 4th day after infection at which time infectivity in the medium was 200 times greater than that associated with the cells. Virus released into the medium was recovered by sedimentation in a sucrose gradient in a continuous-flow centrifuge rotor. Maximal viral infectivity was found at a sucrose concentration of 42%, equivalent to a density of 1.18 g/cm
3
. Deoxyribonucleic acid extracted from these preparations was about 80% viral and 20% cellular as judged by equilibrium centrifugation in cesium chloride density gradients.
Fungal Leaching of Titanium from Rock
Authors: Silverman Melvin P. , Munoz Elaine F.
Published: December 2020

Penicillium simplicissimum
solubilized up to 80% of the titanium in granitic rocks but less than 2% of the titanium in basaltic rocks.
Evaluation of the Modified R/B System for Identification of
Enterobacteriaceae
Authors: Smith P. B. , Tomfohrde K. M. , Rhoden D. L. , Balows A.
Published: December 2020

In comparison with conventional procedures, a modified R/B Enteric Differential System permitted more accurate identification of enteric bacteria than did the original R/B system.
Microbial Esterase Detection with Ultraviolet Fluorescence
Authors: Pancholy Sunil K. , Lynd J. Q.
Published: December 2020

A method is presented to identify esterase-synthesizing microbial colonies within mixed culture plates by utilizing induced esterase hydrolysis of nonfluorescent butyryl ester of 7-hydroxy-4-methylcoumarin to the highly fluorescent 7-hydroxy-4 methyl umbelliferone. Microscopy procedures for making esterase loci of fungal mycelia visible with this reaction are described.
Modification of the Use of the Pipette Filler for Reducing the Hazards of Contamination
Authors: Gioeli R. P. , DeLuca C.
Published: December 2020

An adaptation is described which makes the common propipette readily acceptable for repetitive use. The application of pressure from an appropriate gas to permit rapid delivery of fluids and a method by which the propipette may be used comfortably in one hand make this tool amenable to routine use in a practical attempt to reduce the risks of contamination to both the operator and his cultures.
Comparison of Selective Media for Isolation of Presumptive Group D Streptococci from Human Feces
Authors: Sabbaj Jacobo , Sutter Vera L. , Finegold Sydney M.
Published: December 2020

Pfizer Selective Enterococcus (PSE) agar, a medium containing bile, sodium azide, and esculin, was evaluated for its sensitivity and selectivity for detection and enumeration of presumptive group D streptococci in human feces. SF broth and SF broth plus agar (1.5%), representing selective media in common use, were studied simultaneously. Presumptive group D streptococci were recovered on PSE agar from the feces of all 25 subjects. No growth was observed in 8% of specimens in SF broth. No gram-negative organisms were recovered in any medium. PSE agar has the advantages of selecting out
Streptococcus bovis,
earlier appearance of distinctive reactions, and lack of requirement for special incubation temperature.
Heat Resistance of Spores of Marine and Terrestrial Strains of
Clostridium botulinum
Type C
Authors: Segner W. P. , Schmidt C. F.
Published: December 2020

Resistance to heat of spores of marine and terrestrial strains of
Clostridium botulinum
type C in 0.067
m
phosphate buffer (
p
H 7.0) was determined. The marine strains were 6812, 6813, 6814, and 6816; the terrestrial strains were 468 and 571. The inoculum level equaled 10
6
spores/tube with 10 replicate tubes for each time-temperature variable. Heating times were run at three or more temperatures to permit survival of some fraction of the inoculum. Survivors were recovered at 85 F (30 C) in beef infusion broth containing 1% glucose, 0.10%
l
-cysteine hydrochloride, and 0.14% sodium bicarbonate.
D
values were calculated for each fractional survivor end point after 6 months of incubation. Thermal resistance curves were constructed from the
D
value data.
D
220
(104 C) values for spores of 468 and 571 equaled 0.90 and 0.40 min, respectively. The corresponding values for spores of 6812, 6813, 6814, and 6816 were 0.12, 0.04, 0.02, and 0.08 min. The
z
values for the thermal resistance curves ranged from 9.0 to 11.5 F (5.0 to 6.2 C).
Control of Growth Rate by Initial Substrate Concentration at Values Below Maximum Rate
Authors: Gaudy Anthony F. , Obayashi Alan , Gaudy Elizabeth T.
Published: December 2020

The hyperbolic relationship between specific growth rate, μ, and substrate concentration, proposed by Monod and used since as the basis for the theory of steady-state growth in continuous-flow systems, was tested experimentally in batch cultures. Use of a
Flavobacterium
sp. exhibiting a high saturation constant for growth in glucose minimal medium allowed direct measurement of growth rate and substrate concentration throughout the growth cycle in medium containing a rate-limiting initial concentration of glucose. Specific growth rates were also measured for a wide range of initial glucose concentrations. A plot of specific growth rate versus initial substrate concentration was found to fit the hyperbolic equation. However, the instantaneous relationship between specific growth rate and substrate concentration during growth, which is stated by the equation, was not observed. Well defined exponential growth phases were developed at initial substrate concentrations below that required for support of the maximum exponential growth rate and a constant doubling time was maintained until 50% of the substrate had been used. It is suggested that the external substrate concentration initially present “sets” the specific growth rate by establishing a steady-state internal concentration of substrate, possibly through control of the number of permeation sites.
Effects of Multiple Use on Water Quality of High-Mountain Watersheds: Bacteriological Investigations of Mountain Streams
Authors: Stuart David G. , Bissonnette Gary K. , Goodrich Thomas D. , Walter William G.
Published: December 2020

Bacteriological studies in 1968 and 1969 corroborated earlier findings that a municipal watershed which had been closed to public entry since 1917 yielded water with four to six times the coliform count found in an adjacent mountain watershed open to recreational activities. Similarly, chemical investigations showed higher concentrations of most ions in water from the closed area. Physiological differentiation of coliform and enterococcal bacteria revealed similar types of organisms in both animal droppings and stream water, with fecal coliforms accounting for as much as 70% of the coliform counts observed in the closed area in 1969. Opening of the closed drainage for limited recreation and expanded logging operations in the spring of 1970 coincided with an unexpected decrease in bacterial contamination of that stream. It is postulated that these human activities drove from the watershed a large wild animal population which had contributed substantially to the previous bacterial pollution. It would seem that the practice of closing high-mountain watersheds to public entry is questionable if governmental standards for water quality are to be met, and it also seems that the standards themselves should be reexamined.
Oxidation of Acetate by Various Strains of
Bacillus popilliae
Authors: McKay L. L. , Bhumiratana A. , Costilow Ralph N.
Published: December 2020

A number of strains of
Bacillus popilliae
were examined for their ability to oxidize acetate. Some of these would not sporulate in vitro, and some were oligosporogenous. The ability to oxidize acetate varied widely among the strains tested. A culture derived from spores of the parent strain produced in vivo and one of the asporogenous strains derived from it failed to produce any significant levels of
14
CO
2
from [
14
C]acetate. Oligosporogenous strains derived from the same parent culture all produced
14
CO
2
from both [
1
-
14
C
] and [
2
-
14
C
]acetate but at relatively low rates. The highest rates of acetate oxidation were observed with three strains which did not produce spores in vitro. When cultured under appropriate conditions, one of these strains displayed a secondary growth response concomitant with a decrease in the titratable acidity and an increase in the
p
H of the medium. The data indicate that
B. popilliae
has a complete citric acid cycle but that the activity of the cycle is strongly repressed in wild-type strains under the usual conditions used for in vitro cultivation.
Comparison of Conventional Methods, the R/B System, and Modified R/B System as Guides to the Major Divisions of
Enterobacteriaceae
Authors: Isenberg Henry D. , Painter Barbara G.
Published: December 2020

Primary grouping of more than 2,000 members of
Enterobacteriaceae
, freshly isolated from clinical specimens, was performed on the basis of eight conventional laboratory tests used routinely in this laboratory and by the R/B system. Results indicate that both systems perform well in this first step in the identification of various medically significant species of the family. Difficulties encountered with certain reactions of the R/B system were corrected with a physical modification of the tubes. An additional 100 representatives of
Enterobacteriaceae
were compared with conventional methods and the modified R/B system. Results indicate improvement in observing and interpreting findings with the R/B system.
Effect of Relative Humidity on Formaldehyde Decontamination
Authors: Spiner David R. , Hoffman Robert K.
Published: December 2020

Death rate studies were conducted to determine the effect of varying the concentration, humidity, and type of surface on the sporicidal activity of formaldehyde gas. Washed and unwashed spores were similarly exposed to detect the influence of residual nutrient growth medium upon the rate of kill. The results indicated that the sporicidal activity of formaldehyde gas varies directly with its concentration. Relative humidities (RH) over 50% proved essential for sterility. Spores on a porous surface (cotton cloth) were more readily killed at lower RH than those on a nonporous surface (glass). The reverse occurred at very high RH. At 75% RH, the unwashed spores on glass were killed faster than the washed spores.
Substituted Diazenes: Effect on the Growth of Enterobacteria and Possible Use as Selective Agents for Isolation of Pseudomonads
Authors: Rose Michael J. , Enkiri Nancy K. , Sulzbacher William L.
Published: December 2020

Incorporation of various diazenes into Trypticase soy media appeared selectively to permit the growth of pseudomonads while inhibiting the growth of a variety of enterobacteria. One of these diazenes, diamide (diazenedicarboxylic acid bisdimethylamide), was shown to be bactericidal for pure cultures of
Escherichia coli, Proteus
sp., and
Salmonella enteritidis
and to cause a 1- to 2-hr delay in the growth of
Pseudomonas aeruginosa
. When mixtures of these four organisms were inoculated into Trypticase soy broth or Trypticase soy agar (TSA) containing diamide,
P. aeruginosa
grew in overnight cultures. TSA containing diamide was also used successfully to isolate pseudomonads from soil, clinical urine specimens, fish, ground beef, ground pork, and ground veal.
Observations by Electron Microscopy on Pig Muscle Inoculated and Incubated with
Pseudomonas fragi
Authors: Dutson T. R. , Pearson A. M. , Price J. F. , Spink G. C. , Tarrant P. J. V.
Published: December 2020

Myofibrils from pig muscle inoculated and incubated with
Pseudomonas fragi
showed an extremely disrupted appearance as compared to uninoculated controls. There was an almost complete absence of material in the H zone, marked disruption of the A band (probably myosin), and some loss of dense material from the Z line. These changes indicated that marked proteolysis had occurred. Bacteria observed in spoiled muscle tissue exhibited protrusions or blebs on the outer surface of the cell walls. The blebs appeared to form detached globules that migrated into the muscle mass. Bacteria grown in non-muscle-containing media did not produce blebs, which indicates the blebs were induced by growth on muscle tissue. The possibility that the blebs and globules may contain a proteolytic enzyme responsible for myofibrillar disruption is discussed.
Permeability and Enterotoxic Factors of Nonagglutinable Vibrios
Vibrio alcaligenes
and
Vibrio parahaemolyticus
Authors: Bhattacharya S. , Bose A. K. , Ghosh A. K.
Published: December 2020

Enterotoxic factor (EF) was present and could be developed in nonagglutinable vibrios, but not the permeability factor (PF). Presence of PF without EF in
Vibrio parahaemolyticus
is significant. Both EF and PF were absent in
V. alcaligenes.
Effect of Polycations on Sensitivity of BALB/3T3 Cells to Murine Leukemia and Sarcoma Virus Infectivity
Authors: Manning Jarue S. , Hackett Adeline J. , Darby Norman B.
Published: December 2020

Polybrene or diethylaminoethyl dextran, when added with murine leukemia or sarcoma virus inocula, significantly increased virus infectivity of BALB/3T3 cells. Residual polycation concentrations were nontoxic.
Correlation of Certain Alterations in Metabolic Activity with Alkaloid Production by Submerged
Claviceps
Authors: Řeháček Z. , Sajdl P. , Kozová J. , Malik K. A. , Řičicová A.
Published: December 2020

Alkaloid biosynthesis in
Claviceps paspali
MG-6 was favored by unbalanced growth. A positive correlation between the rate of protein turnover and alkaloid formation was noted. The pattern of the orthophosphate content in the mycelium resembled that in the ripening sclerotia of the parasitic strains. Alkaloids were revealed as potentially effective in energy metabolism. Reduced adenosine triphosphate (ATP) utilization and an increase in the ATP pool were found to be favorable for alkaloid production. Acetyl-coenzyme A carboxylase activity and the level of cell lipids were directly related to the intensity of alkaloid biosynthesis. An inverse relationship was observed between the activities of the tricarboxylic acid and glycoxylate cycles and the rate of alkaloid formation.
Microbial Transformation of Flavonoids
Authors: Ciegler Alex , Lindenfelser Lloyd A. , Nelson George E. N.
Published: December 2020

The ability of a number of fungal spores, and in particular of resting vegetative mycelia, to transform naringin and naringenin was studied. In general, only hydrolytic cleavage of the sugar moieties of naringin to produce prunin and naringenin was observed. Two cultures,
Penicillium charlesii
and
Helminthosporium sativum
, also produced two unidentified flavonoid compounds but in very low yields. No transformation of aglycone was detected, although the compound was metabolized by some cultures when supplied as the glycoside prunin. A fluorodensitometric method was developed for the quantitative analysis of flavonoid compounds.
Enzymatic Production of
l
-Citrulline by
Pseudomonas putida
Authors: Kakimoto Toshio , Shibatani Takeji , Nishimura Noriyuki , Chibata Ichiro
Published: December 2020

To develop an efficient method for the production of
l
-citrulline, optimum conditions for the conversion of
l
-arginine to
l
-citrulline by microbial
l
-arginine deiminase and for production of the enzyme were studied. A number of micro-organisms were screened to test their ability to form and accumulate
l
-citrulline from
l
-arginine.
Pseudomonas putida
was selected as the best organism. With this organism, enzyme activity as high as 9.20 units per ml could be produced by a shaking culture at 30 C in a medium containing glucose, ammonium phosphate,
l
-arginine hydrochloride, yeast extract, peptone, and inorganic salts. Appropriate addition of a surface active agent to the reaction mixture was found to shorten the time required for the conversion. A large amount of
l
-arginine hydrochloride was converted stoichiometrically to
l
-citrulline in 62 hr at 37 C. Accumulated
l
-citrulline was readily isolated in pure form by ordinary procedures with ion-exchange resins. Yields of isolated
l
-citrulline of over 90.5% from
l
-arginine hydrochloride were easily attainable.