bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies on Japanese B Encephalitis Virus Vaccines from Tissue Culture
Authors: Singh Balwant , Hammon W. McD.
Published: December 2020

A study was carried out to evaluate the reliability of and to determine the mechanism involved in an antigen extinction mouse intraperitoneal (ip) challenge test for potency of a cell culture vaccine for Japanese B encephalitis, a modification of a test originated by Sabin for a mouse brain vaccine. Some comparisons were made with the official Japanese test using an intracerebral (ic) challenge after a more prolonged immunization procedure. The Japanese method of using a lyophilized reference vaccine with each test was also employed. It was found that the ip and the ic test appeared to show similar relative differences between lots. The ip test was more quickly and readily performed, gave reasonably consistent results on repetition, and, when used with a suitable reference vaccine, gave promise of being an entirely suitable and reliable test. Immunization by the intramuscular route rather than by the regular ip route appeared to offer no advantage and was less consistent in responses shown. Neutralizing antibody responses of the mice in the standard procedure were very quick to appear, about 4 days after the first dose of vaccine and had a peak titer about the seventh day, the time of challenge. This titer fell quickly unless challenge occurred. The antibody was heat stable, but it was readily inactivated by 2-mercaptoethanol (2-ME). Not until the 11th or 15th day did a small amount of immunoglobulin G appear. Challenge on day 7 significantly increased titers, but this antibody was also mostly inactivated by 2-ME. Interferon did not appear to play any significant role in the protection shown by the mice.
Streptomycin Uptake by
Mycobacterium tuberculosis
Authors: Beggs William H. , Williams Nancy E.
Published: December 2020

Data are presented indicating that
14
C-streptomycin uptake by tubercle bacilli increases as a function of exposure time, is directly and proportionately related to concentration, and involves at least two phases.
Role of Sodium Carboxymethyl Cellulose and Hydroxyethyl Starch in Hematopoietic Cell Line Cultures
Authors: Mizrahi A. , Moore G. E.
Published: December 2020

Sodium carboxymethyl cellulose and hydroxyethyl starch were found to support rapid growth of two hematopoietic cell lines. The polymers were not metabolized by the cells. In the presence of these compounds, lower rates of glucose utilization and lactic acid production were observed. The uptake of glucose by the cells decreased as the concentration of the polymer in the medium was increased. These results indicate that sodium carboxymethyl cellulose and hydroxyethyl starch probably protect the cells against physical stress in suspended cultures.
Biological Activity of the Nucleic Acids Extracted from Two Aerosolized Bacterial Viruses
Authors: Dubovi Edward J.
Published: December 2020

The infectivity of the nucleic acids of πX-174 and MS-2 was examined after aerosolization. It was found that the nucleic acids were undamaged by aerosolization.
Automated Method for the Determination of Free Formalin in Biological Products
Authors: Pemberton John R.
Published: December 2020

A method is described for the rapid and accurate determination of free Formalin in biological products.
Quantitation of Residual Host Protein in Chicken Embryo-Derived Vaccines by Radial Immunodiffusion
Authors: O'Brien Thomas C. , Maloney Clifford J. , Tauraso Nicola M.
Published: December 2020

Quantitation of soluble residual host protein in chicken embryo-derived vaccines was performed rapidly, economically, and accurately by radial immunodiffusion.
Identification of Isolates of
Clostridium perfringens
Types C and D by Agglutination and Fluorescent-Antibody Methods
Authors: Yamagishi Takayoshi , Yoshizawa Jun , Kawai Mitsuteru , Seo Nagaki , Nishida Shoki
Published: December 2020

Agglutination and fluorescent-antibody methods were employed for screening
Clostridium perfringens
types C and D from 393 isolates of this organism. All of 50 strains which were isolated in Japan and were agglutinable with an antiserum prepared against a stock strain of type C no. 3182 toxigenically belonged to type C, but the antiserum showed no cross-agglutination with any of type C strains isolated in Denmark. All of the latter strains, however, were agglutinated by an antiserum prepared against a Danish strain, CWC11. Of 64 strains, showing heat-labile agglutinability by type D antiserum L9, 22 strains were toxigenically identified as type D strains which can be divided into three groups by the heat-stable antigens; no strains which were L-agglutination-positive but O-agglutination-negative were epislon-toxigenic. All of 13 strains, the heat-stable antigen of which was agglutinable by a type D antiserum VX81, were toxigenically type D strains. The results of fluorescent-antibody tests were almost in agreement with those of agglutination test with type C strains and completely with those of the O-agglutination test with type D strains. No beta-, epsilon- or delta-toxigenicity could be demonstrated in strains which were not agglutinated by our test sera for types C and D strains. Further examination of cultural properties of Japanese and Danish type C strains revealed that the two groups were considerably different in urease production, capsule formation, and delta- and alpha-toxigenicities.
Bacteriocin (Marcescin) Typing of Clinical Isolates of
Serratia marcescens
Authors: Traub W. H. , Raymond E. A. , Startsman T. S.
Published: December 2020

A simple, reproducible technique is described for bacteriocin typing of clinical isolates of
Serratia marcescens.
With 10 marcescin-producer strains, a total of 46 of 50 isolates (92%) of
S. marcescens
could be typed and categorized into 16 provisional types according to their sensitivity or tolerance to marcescins. The potential significance of this procedure with regard to delineation of outbreaks of nosocomial infection is discussed on the basis of preliminary epidemiological data.
Enhanced Detection of Australia Antigen in Serum Hepatitis Patients by Discontinuous Counter-Immunoelectrophoresis
Authors: Wallis Craig , Melnick Joseph L.
Published: December 2020

The use of discontinuous counter-immunoelectrophoresis enhanced the reaction between Au/SH antigen and its antibody in agarose. The ionic strength of the Veronal buffer used in the agarose was 0.015 μ, whereas 0.075 μ Veronal (both
p
H 8.6) was used for anode and cathode buffers. Electroendosmosis is increased under such conditions. Au/SH antigen and antibody reacted to give sharp lines within 30 to 45 min as compared with conventional counter-immunoelectrophoresis which required 1 to 3 hr or longer.
Arylamidase of
Cephalosporium acremonium
and Its Specificity for Cephalosporin C
Authors: Dennen David W. , Allen Charles C. , Carver Diane D.
Published: December 2020

Three aggregational forms of arylamidase are produced by
Cephalosporium acremonium
. The exocellular enzyme, with an approximate molecular weight of 60,000, was purified 300-fold by diethylaminoethyl cellulose chromatography, gel filtration, and gel electrophoresis. With
l
-leucyl-β-naphthylamide as the substrate, the
K
m
is 4.2 × 10
−4
m
; the optimum
p
H, 7.7; and the temperature optimum, 35 C. The enzymatic hydrolysis of
l
-leucyl-β-naphthylamide is inhibited by a number of cephalosporins, whereas a variety of penicillins show no effect. Alternatively, the enzyme specifically catalyzes the β-lactam hydrolysis of a number of cephalosporins; a number of penicillins are resistant. The
K
m
for cephalosporin C is 9.09 × 10
−4
m
.
Initiation of Staphylococcal Growth in Laboratory Media
Authors: Genigeorgis Constantin , Martin Sue , Franti Charles E. , Riemann Hans
Published: December 2020

The effects of
p
H and NaCl concentration on the probability of aerobic growth initiation in Brain Heart Infusion broth at 30 C by five staphylococcal strains producing enterotoxins A, B, C, and D were studied in a factorial design experiment. Statistical analysis of the data indicated: (i) significant effects of
p
H, NaCl, and strain on the probability of growth; (ii) diverse effects of NaCl with various
p
H levels and strains; (iii) essentially a linear relationship between NaCl concentration and probability of growth initiation when data for all strains were pooled; (iv) the relationship between NaCl concentration and probability of growth initiation varies from linear to sigmoid, depending on the
p
H of the broth, when the statistically untreated (raw) data are plotted for each strain. Equations were derived which relate the decimal reductions of the number of cells of a staphylococcal population to the concentration of NaCl and
p
H of broth to which the population was exposed. From the equations, the probability that one cell is capable of initiating growth can be calculated. The impact of these types of studies on the development of realistic staphylococcal standards in foods is discussed.
Survival of
Vibrio parahaemolyticus
in Fish Homogenate During Storage at Low Temperatures
Authors: Matches Jack R. , Liston J. , Daneault Louis P.
Published: December 2020

Fish homogenate inoculated with Japanese strains of
Vibrio parahaemolyticus
were either stored at 0.6 C or frozen and stored at −18 and −34 C. Greater survival of the organisms was obtained at 0.6 C than at the lower temperatures.
Assay of Chick Interferons by the Inhibition of Viral Ribonucleic Acid Synthesis
Authors: McWilliams Michael , Finkelstein Martin S. , Allen Patton T. , Giron David J.
Published: December 2020

A method for assaying chick interferons by their inhibition of viral ribonucleic acid synthesis was devised and evaluated. The technique yielded results faster and had more flexibility than other methods with similar sensitivity and reproducibility.
Brine Shrimp (
Artemia salina
L.) Larvae as a Screening System for Fungal Toxins
Authors: Harwig J. , Scott P. M.
Published: December 2020

Concentrations resulting in 50% mortality, determined with brine shrimp (
Artemia salina
L.) larvae exposed to known mycotoxins for 16 hr, were (μg/ml): aflatoxin G
1
, 1.3; diacetoxyscirpenol, 0.47; gliotoxin, 3.5; ochratoxin A, 10.1; and sterigmatocystin, 0.54. 4-Acetamido-4-hydroxy-2-butenoic acid γ-lactone gave no mortality at 10 μg/ml. Used as a screening system involving discs saturated with solutions of known mycotoxins, the larvae were relatively sensitive to aflatoxin B
1
, diacetoxyscirpenol, gliotoxin, kojic acid, ochratoxin A, rubratoxin B, sterigmatocystin, stemphone, and T-2 toxin. Quantities of 0.2 to 2 μg/disc caused detectable mortality. The larvae were only moderately sensitive to citrinin, patulin, penicillic acid, and zearalenone which were detectable at 10 to 20 μg/disc. They were relatively insensitive to griseofulvin, luteoskyrin, oxalic acid, and β-nitropropionic acid. The disc screening method indicated that 27 out of 70 fungal isolates from foods and feeds grown in liquid or solid media produced chloroform-extractable toxic material. Examination of toxic extracts by thin-layer chromatography for 17 known mycotoxins showed that the toxicity of eight isolates could be attributed to aflatoxin B
1
and B
2
, kojic acid, zearalenone, T-2 toxin, or ochratoxin A. Nine out of 32 of these fungal isolates grown in four liquid media yielded toxic culture filtrates from at least one medium. Chemical tests for kojic, oxalic, and β-nitropropionic acids showed the presence of one or two of these compounds in filtrates of seven of these nine isolates.
Production of Ochratoxins in Different Cereal Products by
Aspergillus ochraceus
Authors: Trenk Hugh L. , Butz Mary E. , Chu Fun Sun
Published: December 2020

The effects of temperature and length of incubation on ochratoxin A production in various substrates were studied. The optimal temperature for toxin production by
Aspergillus ochraceus
NRRL-3174 was found to be around 28 C. Very low levels of ochratoxin A are produced in corn, rice, and wheat bran at 4 C. The optimal time for ochratoxin A production depends on the substrate, ranging from 7 to 14 days at 28 C. Ochratoxin B and dihydroisocoumaric acid, i.e., one of the hydrolysis products of ochratoxin A, were produced in rice but at levels considerably lower than ochratoxin A. No ochratoxin C was produced in rice at 28 C. When added to rice cereal or oatmeal, the toxin was found to be very stable over prolonged storage and even to autoclaving for 3 hr.
Rapid Infrared Determination of the Potency of Chlorinated Bactericides
Authors: Spagnolo Frank , Cestaro J. P.
Published: December 2020

A rapid infrared reflectance method for evaluating the germicidal potency of synthetic materials containing various amounts of two chlorinated bactericides was developed. The dimeric product 2,2′-methylenebis (4,6-dichlorophenol) exhibited a characteristic C=C skeletal inplane stretching infrared absorption band at 1,640 cm
−1
. The monomeric 2,4-dichlorophenol precursor showed a characteristic absorption band at 1,579 cm
−1
. These characteristic infrared absorptions may be used for analysis of the potency of the manufactured chlorinated bactericide. For a series of samples known to vary in dimer content, the micrograms per milliliter required for a 100% bacterial kill is first determined by a standard American Petroleum Institute method. Then the area ratio of the infrared absorption bands characteristic of the chlorinated bactericides is measured for each sample and plotted versus the microgram per milliliter required for 100% bacterial kill. The potency of subsequent samples is simply and rapidly determined by measuring this ratio from the infrared absorption curve and calculating micrograms per milliliter required for 100% kill from the calibration curve. Analysis time is approximately 1 hr compared to biocidal tests in current use requiring approximately a 1-month incubation period.
Preparative Electrophoresis of Isotopically Labeled L-Cell Interferons
Authors: Stanček Daniel , Paucker Kurt
Published: December 2020

The preparative method of polyacrylamide gel electrophoresis was adapted for purification and characterization of isotopically labeled L-cell interferons. Re-covery of interferon activity was quantitative, and purification and resolution were comparable to those obtained by analytical polyacrylamide gel electrophoresis. Ultimate specific activities attainable ranged from 2 × 10
6
to 3 × 10
6
international units per mg of protein.
Carbohydrate Fermentation Patterns of
Neisseria meningitidis
Determined by a Microtiter Method
Authors: Davies J. A. , Mitzel J. R. , Beam W. E.
Published: December 2020

A carbohydrate fermentation technique has been developed and compared to the standard fermentation test with cystine-Trypticase-semisolid agar for the identification of
Neisseria meningitidis
. This new method utilizes Mueller-Hinton broth as a basal substrate and is performed with microtiter methods. By using Mueller-Hinton broth with and without the addition of antibiotics, the method can be adjusted to test the fermentation patterns of all of the
Neisseria
including
N. gonorrhoeae
.
Influence of Na
+
on Synthesis of Macromolecules by a Marine Bacterium
Authors: Webb C. D. , Payne W. J.
Published: December 2020

Resting cells of
Vibrio natriegens
acquired the ability to take up
14
C-labeled mannitol in media containing Na
+
and K
+
. But, the cells took up a significant quantity of the label as well in the presence of 0.3
m
K
+
and no Na
+
. The label was distributed throughout the cells in both systems. Cells incubated in mannitol minimal culture medium proliferated and synthesized approximately nine times as much protein in the presence of Na
+
and K
+
as those incubated in the presence of mannitol and 0.3
m
K
+
. The bacteria did not proliferate in the absence of Na
+
. Cells incubated in medium containing mannitol and Na
+
and K
+
synthesized approximately twice the quantity of deoxyribonucleic acid and ribonucleic acid as those incubated in medium containing mannitol and 0.3
m
K
+
but no Na
+
. A significant amount of mannitolbinding protein was synthesized in the membranes of
V. natriegens
incubated in the presence of mannitol and Na
+
and K
+
, but only a small quantity was produced in medium containing mannitol and 0.3
m
K
+
but no Na
+
. A binding fraction comprising at least two proteins (both with molecular weight near 34,000) was isolated by gel electrophoresis from other components of a K
2
CO
3
-extract of membrane protein from mannitol-grown cells. This binding fraction mediated phosphorylation of mannitol at the expense of either adenosine triphosphate or phosphoenolpyruvate. It was then found that mannitol-grown, but not broth-grown, cells contained nicotinamide adenine dinucleotide-linked mannitol-1-phosphate dehydrogenase. Neither contained mannitol dehydrogenase.
Installation and Use of the Microscope Within a Gastight Glove Box
Authors: Boylen Charles W.
Published: December 2020

The adaptation of a high-resolution phase-contrast microscope for glove box use, which allows the stage and focusing knobs in the glovebox, although oculars and camera remain outside, is described.
Self-Sterilizing Inoculating Loop
Authors: Songer J. R. , Riley J. L. , Braymen D. T.
Published: December 2020

A self-sterilizing inoculating loop consisting of a step-down transformer and an adjustable timing circuit are described.
Antigenic Analysis of
Rhizobium japonicum
by Immunodiffusion
Authors: Dudman W. F.
Published: December 2020

Immunodiffusion reactions were studied with seven strains of
Rhizobium japonicum
and three strains of the cowpea miscellany by using antisera against eight of the strains. Most strains yielded only weak precipitin bands when untreated cell suspensions were used as antigens in the diffusions. Ultrasonic disruption or heat treatment of the cells led to stronger bands, and immersion in boiling water for 20 min was used as the standard procedure for preparing these bacteria for immunodiffusion analysis. Heat-labile antigens were detected in only a few strains; the major antigens of all of the strains appeared to be heat-stable. Many of the strains cross-reacted, sometimes in a nonreciprocal manner; unheated cell suspensions cross-reacted more widely but more weakly than the heated suspensions. Heat-treated crushed nodule preparations reacted well in immunodiffusions. The antigens of cultured cell and nodule extract (bacteroid) forms of three strains were compared. In one of these strains, an antigen present in the cultured cells was absent from the bacteroids. Unknown strains present in soybean root nodules were readily identified by immunodiffusion.
Growth and Cultivation of the Unusual Generalized Transducing
Bacillus
Bacteriophage SP-15
Authors: Taylor Martha J. , Goldberg Ivan D.
Published: December 2020

Additional properties of SP-15, a generalized transducing bacteriophage notable for the ability to transfer an unusually large fragment of deoxyribonucleic acid (DNA) to
Bacillus subtilis
and
B. licheniformis
, are presented together with improved methods that enhance its utility. Simple means have been found to provide the rigid control over moisture that is necessary for the assay of plaque-forming units (PFU). Reproducible procedures for propagating transducing phage, which depend upon an appropriate mixing of PFU with uninfected bacteria, have replaced less reliable methods that utilized infected spores. Transduction of
B. subtilis
W-23 increased linearly when MgSO
4
in recipient cell-SP-15 mixtures was increased from 0.005 to 0.03
m.
Methods have been developed that protect SP-15 from the damaging effects of CsCl and of osmotic shock subsequent to dilution. Evidence that the PFU and transducing particles of lysates decay at the same slow rate during extended storage suggests that the decay is a result of damage to protein rather than to DNA. One-step growth experiments, in which SP-15 was propagated on
B. subtilis
W-23-S
r
/1 mg, indicated a latent period of 100 min, a rise period of 60 min, and a burst size of 25 to 34 PFU per infected cell. These findings suggest explanations for some of the technical difficulties SP-15 has presented.
Effect of the Method of Agitation on the Accumulation of Protective Antigen in Cultures of
Bacillus anthracis
Authors: Wright George G. , Angelety Lorell H.
Published: December 2020

Accumulation of protective antigen was essentially independent of culture volume, provided the organisms were kept in suspension by stirring. Agitation by orbited shaking was unsatisfactory.
Rapid Detection of Viral Antibody by Cellulose Acetate Electrophoresis
Authors: Levitt Neil H. , Amsler Kenneth R. , McKinney Robert W.
Published: December 2020

An immunoelectrophoretic procedure utilizing microprecipitation and cellulose acetate electrophoresis was developed for detection of antibody to specific virus. A model system of tobacco mosaic virus and homologous rabbit antiserum is described.
Antigens of
Pasteurella tularensis
: Preparative Procedures
Authors: Nutter John E.
Published: December 2020

Ether-water (EW) extraction of
Pasteurella tularensis
produced better antigens than five other chemical procedures. EW extracts produced from stationary-phase, liquid-grown, saline suspensions of strain SCHU S4 cells regularly induced agglutinin and precipitin formation in rabbits. Mice, guinea pigs, and monkeys also responded to EW extracts but with lower antibody levels. The use of strains of lower virulence, acetone-dried cells, organisms grown on a solid medium, and abbreviated extraction conditions all resulted in extracts with a diminished antigenicity, but logarithmic-phase and stationary-phase cells yielded equivalent EW extracts. The use of adjuvant, hyperimmunization, and large doses of antigen increased the precipitin responses of rabbits without appreciably altering the agglutinin response. By the appropriate combination of centrifugal fractionation of EW extracts, use of adjuvant, and vaccination schedule, rabbit antisera with either predominantly agglutinating or precipitating activities were obtained.
Biotin Production and Utilization in a Sewage Treatment Lagoon
Authors: Fillipi Gordon M. , Vennes John W.
Published: December 2020

Biotin, in a sewage oxidation lagoon also receiving potato processing wastes, was observed to increase two logs during the summer period of waste stabilization and then to decline to near earlier concentrations. Three organisms,
Aerobacter aerogenes, Chlorella vulgaris
, and
Thiocapsa floridana
, were at least partially responsible for these fluctuations; the latter two organisms were associated with biotin utilization and the former with biotin production. Since copious quantities of biotin are produced in these waste treatment facilities, the vitamin may act as a useful indicator of microbial action on certain organic molecules, especially in domestic and industrial wastes such as those from municipalities and potato and sugar beet processing plants. Furthermore, the presence of biotin in rivers and streams may be indicative of the discharge of incompletely stabilized wastes from these sources.
In Vitro Sensitivity of
Torulopsis glabrata
to Amphotericin B, 5-Fluorocytosine, and Clotrimazole (Bay 5097)
Authors: Marks Melvin I. , Steer Paul , Eickhoff Theodore C.
Published: December 2020

Thirty-five strains of
Torulopsis glabrata
were tested by a tube dilution method for their susceptibility to amphotericin B, 5-fluorocytosine, and clotrimazole (Bay 5097). Amphotericin B was the most active in vitro, inhibiting all strains at a concentration of 1 μg/ml and killing all strains at 2 μg/ml. 5-Fluorocytosine inhibited over 80% of strains at 0.24 μg/ml, but three strains required ≥7.8 μg/ml for killing. A concentration of 2 μg of clotrimazole per ml inhibited less than 50% of strains, and 8 μg/ml killed only 10% of strains. Most strains of
T. glabrata
were killed by therapeutically achievable concentrations of amphotericin B and 5-fluorocytosine, but not clotrimazole.
Beta Hemolysin: a Persistent Impurity in Preparations of Staphylococcal Nuclease and Enterotoxin
Authors: Chesbro William , Kucic Vera
Published: December 2020

Purified staphylococcal nuclease and enterotoxin B from several sources contained beta hemolysin whose physicochemical resemblances to the other two proteins make its elimination difficult.
Effect of Serum on the Yield of Lymphocytic Choriomeningitis Virus in Baby Hamster Kidney Cells
Authors: Stanwick Trevor L. , Kirk B. E.
Published: December 2020

The yields of the Armstrong and WE strains of lymphocytic choriomeningitis virus in baby hamster kidney (BHK) cells cultivated in either bovine, calf, fetal bovine, or horse serum were investigated. Lines of BHK cells were established in these sera. When the infected cell lines were observed by immunofluorescence, the per cent fluorescing cells for a given virus strain did not vary. However, for both strains, the extracellular virus yields per cell were significantly greater in the fetal bovine-cell line than in the other serum-cell lines.
In Vitro Chemotherapeutic Combinations Against Isoniazid-Resistant
Mycobacterium tuberculosis
and
Mycobacterium fortuitum
Authors: Goldstein Elliot , Eagle M. Carroll , LaCasse M. L.
Published: December 2020

It is an acceptable medical practice to use second-line antimycobacterial drugs in combination with isoniazid in treatment of isoniazid-resistant tuberculosis. Recent investigations have demonstrated the importance of determining chemotherapeutic interaction in instances of multiple antibiotic use. We studied the inhibitory effect of combinations of isoniazid with ethambutol, rifampin, ethionamide, cycloserine, viomycin, and kanamycin against three isoniazid-resistant strains of
Mycobacterium tuberculosis
and three strains of
M. fortuitum.
The isobologram technique with drug concentrations of 0.4 to 100 μg/ml was used. With the exception of single instances in which kanamycin plus isoniazid (
M. tuberculosis
strain 9999) and ethionamide plus isoniazid (
M. fortuitum
strain 2080) seemed to have a synergistic effect, neither synergy nor antagonism was noted for any of the combinations. These studies show that the combined use of isoniazid and a second line antimycobacterial agent results in vitro in indifferent inhibitory activity.
Efficacy and Safety of Topical Lysostaphin Treatment of Persistent Nasal Carriage of
Staphylococcus aureus
Authors: Quickel Kenneth E. , Selden Richard , Caldwell Jacques R. , Nora Nedo F. , Schaffner William
Published: December 2020

The efficacy of lysostaphin nasal spray and Neosporin ointment (Burroughs Wellcome & Co.) in altering nasal carriage of
Staphylococcus aureus
was studied with persistent carriers in an institution for mentally retarded children and adults. Treatment for 5 days with either agent significantly reduced carriage rates. This effect persisted through the 5th day after therapy with lysostaphin but not with Neosporin. By the 11th day after therapy, carriage rates in the treatment and control groups were not significantly different. Except for a single immediate wheal and flair skin test reaction, no other evidence of adverse reactions to topical lysostaphin was detected. No consistent changes in hemagglutination-inhibition titers to lysostaphin were observed after therapy. Lysostaphin appears to be slightly more effective than conventional topical antimicrobial therapy in reducing nasal carriage of staphylococci in this rigorously defined population of persistent carriers.
Determination of Fungal Elastase
Authors: Rippon John W. , Hoo Martin Soo
Published: December 2020

The elastin-agar plate is useful for determining elastolytic activity. Determination of fungal elastase, however, requires partial purification of culture extracts and the orcein elastin or gravimetric method.
Control of Contamination in Psittacosis Specimens by Antibiotics
Authors: Lewis Vester J. , Engelman Helen M.
Published: December 2020

Diluent containing a combination of vancomycin, kanamycin, and streptomycin is shown to be more effective than diluent previously used for isolating
Chlamydia psittaci
from bird tissue specimens.
Pigment Production of
Cryptococcus neoformans
Grown with Extracts of
Guizotia abyssinica
Authors: Strachan A. A. , Yu R. J. , Blank F.
Published: December 2020

Brown pigment(s) formed in
Cryptococcus neoformans
when grown on media containing extracts of the seeds of
Guizotia abyssinica
cannot be extracted by common organic solvents or by 6
n
HCl or 2
n
NaOH. A similar pigmentation was observed in
C. neoformans
when grown on a medium containing caffeic acid isolated from the hydrolyzed methanol extract of
G. abyssinica
seeds. Its methyl ester and the diacetate thereof, as well as the following structurally related compounds, 3-hydroxytyramine, 3,4-dihydroxybenzoic acid, 3,4-dihydroxyphenylethanolamine, and 4-hydroxy-3,5-dimethoxycinnamic acid, brought about similar pigmentation. However, 2,4-, 2,5-, 2,6-, and 3,5-dihydroxybenzoic acids, tyrosine, phenylalanine, cinnamic acid, 4-hydroxycinnamic acid, and 4-hydroxy-3-methoxycinnamic acid did not cause coloration in
C. neoformans.
Formation of Aerial Hyphae in
Candida albicans
Authors: Iralu Vichazelhu
Published: December 2020

Each of 22 isolates of
Candida albicans
was induced to produce aerial hyphae by culturing on a solid medium containing a peptone, acid-hydrolyzed casein, soluble starch, and agar in an atmosphere of 10% CO
2
at 37 C and room temperatures. Production of aerial hyphae is not diagnostic for
C. albicans.
Some of the other species of
Candida
may also produce such structures.
Incidence of Prosthecate Bacteria in a Polluted Stream
Authors: Staley James T.
Published: December 2020

Water samples were collected aseptically several times throughout the year at nine stations on the Red Cedar River, a stream flowing through farmland and receiving effluent from several municipalities in central Michigan. Total prosthecate bacteria were enumerated by both direct and viable counting techniques. By direct techniques, these bacteria accounted for 0.62 to 1.1% of the total microflora during the study. The predominant type of appendaged bacteria was the caulobacters (
Caulobacter, Asticcacaulis
, and the fusiform caulobacter), which accounted for 64 to 93% of the total prosthecate forms. The others of importance were prosthecomicrobia (< 1 to 24%), including
Prosthecomicrobium and Prosthecochloris
; hyphomicrobia (< 1 to 15%), including
Hyphomicrobium
and
Rhodomicrobium
; and
Ancalomicrobium
(< 1 to 6%). The viable counts of heterotrophs also indicated that the caulobacters were the most numerous prosthecate bacteria in the stream. They ranged from fewer than 1 per ml to a maximum of almost 4,000 per ml. During the coldest period, when the total viable counts decreased to about 10
4
per ml compared to their summer high of over 10
7
per ml, the caulobacters actually increased in numbers. In December (temperature 0 to 1 C), they comprised from 0.09 to 1.0% of the viable microbial count, and in March (6.0 to 8.0 C) they accounted for 0.14 to 2.8%. The other heterotrophic prosthecate bacteria were generally found at numbers less than 1 per ml, with the exception of the December study when
Hyphomicrobium
was present in numbers as high as 2,400 per ml. There was no consistent correlation between the frequency of prosthecate bacteria and total coliforms in the stream during the investigation.
Bleomycin, an Antitumor Antibiotic: Improved Microbiological Assay and Tissue Distribution Studies in Normal Mice
Authors: Pittillo Robert F. , Woolley Carolyn , Rice Louise S.
Published: December 2020

A microbiological assay was developed for bleomycin, an antitumor antibiotic reported to be active in human trials. The assay bacterium was a strain of
Escherichia coli
which is resistant to ethionine. Studies revealed relatively high concentrations of bleomycin in the blood and urine of mice after a single dose, < 0.33
ld
10
, injected intraperitoneally.
Enumeration and Identification of
Bacillus cereus
in Foods
Authors: Kim H. U. , Goepfert J. M.
Published: December 2020

An egg yolk-polymyxin medium (KG) for rapid enumeration of
Bacillus cereus
is described. The test is presumptive in that differentiation of
B. cereus
(and closely related organisms) from other species is based on the formation of turbidity in the agar surrounding the colonies of the
cereus
group organisms. The medium is formulated to encourage sporulation and release of free spores for serological confirmatory tests within the 24-hr incubation period. The production of turbidity in egg yolk and free-spore production by 25 strains of
B. cereus
on KG agar were measured. The recovery of food poisoning strains of
B. cereus
inoculated into nonsterile food slurries was assessed. A comparison of KG agar and mannitol-egg yolk-polymyxin-agar indicated that the two media were comparable in their abilities to recover low levels of
B. cereus
from naturally contaminated foods. Since KG agar enhances spore formation by
B. cereus
, thus permitting early serological testing, its use in screening food products is advocated.
Recovery of Small Quantities of Viruses from Clean Waters on Cellulose Nitrate Membrane Filters
Authors: Berg Gerald , Dahling Daniel R. , Berman Donald
Published: December 2020

A method is described for quantitatively recovering small amounts of viruses from large volumes of buffered, distilled water. Development of the method was motivated by the anticipated need for testing large volumes of renovated sewage for viruses. The method consists of adsorbing viruses onto cellulose nitrate membrane filters (0.45 μm pore size) from water containing sufficient Na
2
HPO
4
to produce a molarity of 0.05 and sufficient citric acid to produce a
p
H of 7, and eluting the adsorbed viruses in 3% beef extract under extended sonic treatment. Complete recovery of poliovirus 1, echovirus 7, and coxsackievirus B3 resulted when less than 100 plaque-forming units were added to 1-liter quantities of water. Recoveries of reovirus 1 were almost as good. Preliminary studies indicate that good recoveries can be made from 25-gal quantities of water. The method described is efficient in waters of high quality and may be useful for recovering viruses in renovated, and perhaps in tap waters, but not in waters containing certain organic matter unless that matter is first removed.
Effect of β-Propiolactone on Sendai Virus
Authors: Wainberg M. A. , Hjorth R. N. , Howe C.
Published: December 2020

The biological properties (infectivity, hemagglutination, hemolysis, cell fusion, neuraminidase) of Sendai virus were dissociated on the basis of sensitivity to β-propiolactone, by freeze-thawing, by heating at different temperatures, and by adsorption-elution with formalinized chicken erythrocytes. Possible mechanisms whereby β-propiolactone selectively destroys viral infectivity are discussed.
Comparison of Schaedler Agar and Trypticase Soy-Yeast Extract Agar for the Cultivation of Anaerobic Bacteria
Authors: Starr S. E. , Killgore G. E. , Dowell V. R.
Published: December 2020

Schaedler agar (SA) and Trypticase soy-yeast extract agar (TSYEA), both supplemented with rabbit blood (5%, v/v) and menadione (0.5 mg/liter), were compared with respect to quantitative recovery, quality of growth, and rapidity of growth of selected anaerobic bacteria. The media were stored for 2 to 4 days prior to use in an anaerobic glove box, where all subsequent bacteriological procedures were performed. After 24 hr of incubation, colonies of
Clostridium cadaveris (C. capitovale), C. haemolyticum, C. novyi
A, and
C. perfringens
were larger on SA than on TSYEA, and the appearance of
C. novyi
B colonies on SA at 24 hr antedated their appearance on TSYEA. Quantitative recovery of
C. novyi
B was improved on SA; recovery of the other clostridia tested was comparable on the two media (inconclusive results were obtained with
C. novyi
A). Rough colonial types of some of the clostridia emerged on SA. No appreciable differences in results with the two media were noted for
Bacteroides fragilis, B. melaninogenicus,
or
Fusobacterium fusiforme.
Improved Medium for Enumeration of
Clostridium perfringens
Authors: Harmon Stanley M. , Kautter Donald A. , Peeler James T.
Published: December 2020

An improved selective medium, Tryptose-sulfite-cycloserine (TSC) agar, for the enumeration of
Clostridium perfringens
is described. It consists of the same basal medium as Shahidi-Ferguson-perfringens (SFP) agar, but with 400 μg of D-cycloserine per ml substituted for the kanamycin and polymyxin. Tolerance of
C. perfringens
for D-cycloserine, its production of lecithinase, and its ability to reduce sulfite were used as the basis for development of this medium. Comparisons were made between TSC and SFP agars for the recovery of vegetative cells of
C. perfringens
by using statistical methods. The results showed that TSC allowed virtually complete recovery of most of the
C. perfringens
strains while inhibiting practically all facultative anaerobes tested. SFP agar allowed a slightly higher rate of recovery of
C. perfringens
but was found to be much less selective.
Characteristics of Antimicrobial Resistance of
Escherichia coli
from Animals: Relationship to Veterinary and Management Uses of Antimicrobial Agents
Authors: Mercer H. D. , Pocurull D. , Gaines S. , Wilson S. , Bennett J. V.
Published: December 2020

Five-hundred fifty-five (555) isolates of
Escherichia coli
were obtained from fecal specimens of a representative number of animals from five farms in the United States. Antibiotic exposure of the selected herds was determined by an epidemiological survey of these farms. The incidence of multiple resistance in the
E. coli
isolates was higher in herds exposed to continuous feeding of antimicrobial agents (84.8%) than in a herd not receiving antimicrobials (15.7%). The most common resistance configuration observed was the triple pattern of dihydrostreptomycin (DS), sulfonamide (SU), and tetracycline (TE). The second most frequent pattern consisted of four resistances: ampicillin (AM), DS, SU, and TE. The frequency of transfer factors was much higher in multiply resistant organisms from the herds exposed to antimicrobial medicaments. The
E. coli
isolates were relatively efficient in fostering and transferring heterologous resistance factors. AM resistance factors occurred more frequently in herds which were exposed to feed levels of penicillin (27.9%) than in those that were not (6.4%).
Rapid Quantitative Microbiological Assay of Antibiotics and Chemical Preservatives of a Nonantibiotic Nature
Authors: Kabay Antal
Published: December 2020

A rapid quantitative assay of 14 antibiotics and 6 chemical preservatives by the cylinder-plate diffusion method is described, in which
Bacillus stearothermophilus
ATCC 12980 was used as the test organism. After 4 hr of incubation at 65 C, the zones of inhibition can be read off easily with the naked eye. The 4-hr assay was compared under identical conditions with the conventional 16- to 18-hr agar diffusion assay, i.e., by using the test organisms and media proposed by the
Code of Federal Regulations
, and no difference in accuracy between the two methods was detected. In both cases, the coefficient of variation for replicate tests was less than 2%. The test strain is highly sensitive to penicillins, cephalosporins, and bacitracin and is consequently also particularly suitable for the determination of low-level concentrations of these substances in aqueous solutions and in urine.
Detection of Bacteremia in Children with Sodium Polyanethol Sulfonate: a Prospective Clinical Study
Authors: Minkus Robert , Moffet Hugh L.
Published: December 2020

A prospective study was made of 1,000 consecutive duplicate blood cultures obtained from sick children to evaluate the usefulness of sodium polyanethol sulfonate (SPS). With the small volumes of blood (1 to 5 ml) usually obtained for blood cultures in children, SPS did not increase the frequency of recovery of organisms judged to be associated with clinical infections, with the possible exception of
Diplococcus pneumoniae.
However, the use of SPS was associated with an increased frequency of recovery of organisms judged to be contaminants, such as
Staphylococcus epidermidis
and propionibacteria, possibly because SPS enhanced the recovery of a very small inoculum of skin bacteria.
Anaerobic CO
2
Cabinet for the Cultivation of Strict Anaerobes
Authors: Leach P. Ann , Bullen J. J. , Grant I. D.
Published: December 2020

The design and construction of an anaerobic CO
2
cabinet are described. Air is displaced by a stream of oxygen-free CO
2
, and anerobic conditions are produced in 3 hr. The equipment is simple and cheap to operate and has been found to be satisfactory for the isolation of strict anaerobes from the mouse intestine.
Inactivated Eastern Equine Encephalomyelitis Vaccines Prepared in Monolayer and Concentrated Suspension Chick Embryo Cultures
Authors: White A. , Berman S. , Lowenthal J. P.
Published: December 2020

Eastern equine encephalomyelitis vaccines were prepared with virus propagated in stationary monolayer cultures and in concentrated suspension cultures of primary chick embryo cells. Virus pools for vaccine preparation were inactivated by three different methods: 0.05% formalin, 41 C heat, and 0.16% β-propiolactone. Heat-and β-propiolactone-inactivated vaccines maintained high hemagglutinating titers in the fluid state for at least 10 months, whereas formalin-inactivated vaccines lost their hemagglutinating activity within a few hours after treatment. The hemagglutinin of β-propiolactone-inactivated virus particles was more dense than the hemagglutinin of the parent virus particles, as determined by sucrose density gradient centrifugation. The increase in density may be due to the degradation or removal of the lipid from the virus envelope. When administered to guinea pigs, all three vaccines stimulated hemagglutination-inhibiting, complement-fixing, and neutralizing antibodies and afforded protection against a live virus challenge. Test results showed that vaccines prepared with virus propagated in concentrated suspension cultures were more immunogenic and stimulated greater serologic responses in guinea pigs than vaccines derived from monolayer-propagated virus. The β-propiolactone-inactivated vaccine was most protective, the heat-inactivated (41 C) vaccine next, and the formalin-inactivated vaccine least potent.
Preparation of Type 2 Herpes Simplex Virus Complement-Fixing Antigen
Authors: Palmer Erskine L. , Martin Mary Lane , Warfield Donna T.
Published: December 2020

Comparable complement-fixing antigens of type 1 and type 2 herpes simplex virus were produced by extraction of infected African green monkey cells with 0.85% NaCl which was buffered at
p
H 9.0 with 0.05
m
glycine-NaOH. The optimal antigen dilutions were higher in titrations against hyperimmune animal sera than in titrations against human sera. Complement-fixing antibody to type 2 herpes antigen was detected in 5 of 17 sera from healthy humans.
Procedure for the Transfer of Small Discs of Agar-Containing Media Under Sterile Conditions and Some Applications of This Technique (Agar Disc Auxanography)
Authors: Woolfolk C. A.
Published: December 2020

Small agar discs are formed and transferred under sterile conditions. This technique permits the efficient and versatile use of the petri plate when many growth conditions and (or) cultures are being examined.
Bacteriophage Types and Antibiotic Susceptibility of
Staphylococcus aureus
Authors: Klastersky J. , Beumer J. , Daneau D.
Published: December 2020

In vitro tests of 132 strains of
Staphylococcus aureus
, among which were 38 (28.7%) heteroresistant strains, were performed with 14 commonly used antibiotics, including gentamicin and vancomycin. Heteroresistant strains were found predominantly, but not only, with group 3 phage-typable strains; no heteroresistant strain was found in out-patients. Gentamicin appeared as a uniformly effective agent at low concentrations against oxacillin-sensitive and -resistant strains. It is suggested that gentamicin should be compared to vancomycin in future clinical therapeutic trials in severe staphylococcal infections, especially in cases due to oxacillin-resistant strains.
Physiological Studies of an Oligosporogenous Strain of
Bacillus popilliae
Authors: Costilow Ralph N. , Coulter Wilson H.
Published: December 2020

A relatively small but consistent increase in the frequency of spore formation by an oligosporogenous strain of
Bacillus popilliae
(NRRL B-2309M) was obtained by adding 0.1% sodium pyruvate to the sporulation medium. The frequency of spore formation was essentially the same when a low level of glucose, trehalose, or glucose-6-phosphate or a high level of α-methyl-
d
-mannoside was added as the carbon and energy source. Many other variations in the cultural medium and cultural conditions failed to enhance spore formation of 2309M, and no spores were found in four asporogenic strains under any of the conditions tried. There were no significant differences between the 2309M strain and three nonsporeforming cultures with respect to (i) the rate and extent of growth, (ii) the rates of glucose utilization, or (iii) volatile acid production and utilization. None of the cultures tested was found to produce detectable levels of extracellular protease or an antibiotic. The only consistent marker found associated with spore formation was the development of catalase activity, and this activity was stimulated by heating at 80 C for 10 min. This was not found unless morphological evidence of spore formation was observed. The germination of the spores formed by 2309M in vitro was stimulated by heat shock and by the addition of pyruvate to the germination medium.
Metabolism of Pyrimidine Nucleotides in a Microorganism
Authors: Sakai Takuo , Watanabe Taizo , Chibata Ichiro
Published: December 2020

A study was made to develop a new method for the production of ribose-5-phosphate (R-5-P) from uridine-5′-monophosphate (UMP) by the action of nucleotide-
N
-ribosidase of
Pseudomonas oleovorans
, and a suitable medium for the formation of nucleotide-
N
-ribosidase was established. For the enzymatic conversion of UMP to R-5-P, a cell suspension was employed as the enzyme source. Although degradation of R-5-P, the desired product, occurred during the course of the enzyme reaction, it was prevented by the addition of an appropriate amount of zinc ion and resulted in a stoichiometric conversion of UMP to R-5-P and uracil. Accumulated R-5-P was readily isolated by ion-exchange chromatography of the bacteria-free reaction mixture. Yield of isolated R-5-P was about 60% of the theoretical recovery.
Lysine Decarboxylase Activity in Broth and Agar Media
Authors: Elston Harry R.
Published: December 2020

Four lysine decarboxylase media were studied by testing them with 305
Enterobacteriaceae
and 42 nonfermenting bacilli. A comparison was made between lysine decarboxylase broth medium (Moeller base) and Johnson's semisolid agar without lactose and Bachrach's broth medium and lysine-agar slants which contain lactose. The nonlactose media, lysine decarboxylase broth and the semisolid medium of Johnson, were the best media for use with all of the bacteria studied. The exclusion of lactose from lysine decarboxylase medium seems desirable to extend the usefulness of this medium among members of the
Enterobacteriaceae.
When the results with lysine decarboxylase broth and Johnson's semisolid medium without lactose were compared, a 6% difference existed between the results obtained with lysine decarboxylase broth and Johnson's semisolid agar. When the results with Bachrach's broth and lysine-agar slants with lactose were compared, a 1% difference existed between Bachrach's broth and the agar slant method. At times, reading and interpretation were difficult because of intermediate degrees of color change. The inability of
Pseudomonas aeruginosa
or
Herellea
to utilize glucose under the anaerobic condition of the medium makes the lysine decarboxylase test an undesirable procedure for these organisms. Of the four test media used, the lysine-lactose-agar slants seemed to be the least desirable because of the more frequent occurrence of indistinct color reactions and shifts in color.