bluuebunny/crossref_metadata_2025 huggingface dataset:
Gelatin-induced Reversion of Protoplasts of
Bacillus subtilis
to the Bacillary Form: Biosynthesis of Macromolecules and Wall During Successive Steps
Authors: Landman Otto E. , Forman Arnold
Published: January 2020

Protoplasts of
Bacillus subtilis
plated on SDG medium formed L colonies in quantative yield and propagated in the L-form indefinitely. Protoplasts or L bodies placed in 25% gelatin medium formed bacillary colonies. Details of the reversion of these naked bodies to the walled form are reported here. Protoplasts prepared in minimal medium reverted fairly synchronously 3 to 4 hr after inoculation into gelatin, but protoplasts preincubated in casein hydrolysate (CH)-enriched minimal medium were primed to revert within 1 hr in the gelatin. Preincubation for 1.5 hr in 0.44% CH was required for good priming. Cells must be subjected to this preincubation (step 1) in the naked state; it is effective for L bodies as well as protoplasts. Priming was blocked by chloramphenicol, puromycin, and actinomycin D but was not affected by penicillin, lysozyme, or inhibition of deoxyribonucleic acid (DNA) synthesis. It is concluded that protein and ribonucleic acid (RNA) synthesis are required during step 1, that DNA synthesis is not required, and that wall mucopeptide is not made. The reversion of well-primed protoplasts in the gelatin (step 2) proceeded undisturbed in thymine-starved cells with chromosomes arrested at the terminus. It was scarcely slowed by chloramphenicol in the gelatin but was delayed about 3 hr by both puromycin and actinomycin D. Escape from inhibition occurred while the inhibitors were still actively blocking growth. Penicillin and cycloserine inhibited and lysozyme reversed reversion. Momentary melting of the gelatin delayed reversion. It is concluded that mucopeptide synthesis occurs in step 2, that concomitant RNA, DNA, or protein synthesis is not essential, but that physical immobilization of excreted cell products at the protoplast surface is necessary early in step 2. Newly reverted cells were misshapen and osmotically sensitive. Processes which confer osmotic stability after reversion (step 3) did not occur in the presence of chloramphenicol or actinomycin D.
Rhodopseudomonas acidophila
, sp. n., a New Species of the Budding Purple Nonsulfur Bacteria
Authors: Pfennig Norbert
Published: January 2020

A succinate-mineral salts medium of
p
H 5.2 provided selective enrichment conditions for
Rhodomicrobium vannielii
and for a new species belonging to the
Athiorhodaceae
, described herein as
Rhodopseudomonas acidophila
. Seven strains of the new species have been isolated from different sources in the United States and Germany. The cells are rod-shaped or ovoid, 1.0 to 1.3 μm wide and 2 to 5 μm long, and motile by means of polar flagella. Multiplication occurs by budding. The photopigments consist of bacteriochlorophyll
a
and carotenoids of the spirilloxanthin series, together with new carotenoids. All strains can grow either under anaerobic conditions in the light or under microaerophilic to aerobic conditions in the dark. No growth factors are required. The range of simple organic substrates photo-assimilated resembles that characteristic of
Rhodomicrobium
. Good photolithotrophic growth is possible at the expense of molecular hydrogen; thiosulfate and sulfide are not utilized.
Ultrastructure of
Brucella abortus
L-Forms Induced by Penicillin in a Liquid and in a Semisolid Medium
Authors: Hatten Betty A. , Schulze Mary L. , Huang Shyi Yi , Sulkin S. Edward
Published: January 2020

Brucella abortus
L-forms were induced by 5.0 or 10.0 μg of penicillin/ml in a broth medium containing 0.3
m
sucrose, and in a semisolid medium containing 10% calf serum and 20.0, 40.0, or 60.0 μg of penicillin/ml. After 96 hr of incubation, L-forms of various sizes and shapes were observed. Basic structures of the L-forms were similar whether induced in liquid or semisolid medium. L-forms had two “unit” membranes, each consisting of two outer dense layers separated by a lucent layer. A few large, irregularly shaped organisms in penicillin-treated broth cultures had additional surface material and were referred to as “transitional” forms. In contrast with L-forms, the bacterial cells were fairly uniform in size and shape, were smaller, and had a more complex cell wall structure. Small bodies limited by a “unit” membrane were present within and around numerous L-forms from liquid and semisolid medium cultures. Other internal membranous structures were also seen in some L-forms. Most
Brucella
L-forms described in this paper reverted to bacteria in the absence of penicillin and were structurally characteristic of unstable L-forms.
Rhodospirillum tenue
sp. n., a New Species of the Purple Nonsulfur Bacteria
Authors: Pfennig Norbert
Published: January 2020

Rhodospirillum tenue
sp. n. differs from the rhodospirilla described so far by its small size, ultrastructure, and physiological and biochemical characteristics.
Induced Bactericidal Response in the Hagfish
Authors: Acton Ronald T. , Weinheimer Peter F. , Hildemann W. H. , Evans E. Edward
Published: January 2020

Hagfish were shown to be capable of synthesizing bactericidins after injection of gram-negative bacteria. The bactericidins could be detected as early as 2 days after injection. The degree of specificity was not as impressive as in mammalian systems.
Lymphogranuloma Venereum II. Characterization of Some Recently Isolated Strains
Authors: Schachter Julius , Meyer K. F.
Published: January 2020

Five Bedsonia (Chlamydia) isolates from lymphogranuloma venereum (LGV) patients were tested for inclusion type, sulfonamide sensitivity, and mouse virulence. Two matched the classical description of LGV agents. Two were not virulent for mice by the intracerebral route, therefore fitting the description for trachoma-inclusion conjunctivitis agents. One was highly virulent for mice and sulfonamide-resistant, and produced inclusions that did not stain with iodine, all characteristics generally associated with avian bedsoniae. A sixth isolate could not be adequately tested due to poor infective yields. Because of this variety of properties within the Bedsonia group, the term LGV might more appropriately be reserved for clinical disease rather than to describe a particular bedsonial agent.
Somatic O Antigen Relationship of
Brucella
and
Vibrio cholerae
Authors: Feeley John C.
Published: January 2020

The antigenic relationship between
Brucella
species and
Vibrio cholerae
was examined by agglutinin and agglutinin-absorption tests by using rabbit antisera. Brucella antisera agglutinated only the Inaba serotype of
V. cholerae
and at low titer. Inaba-reactive antibody was absorbed by either heat-stable (100 C, 2 hr) Ogawa or Inaba O antigens. Cholera antisera from rabbits immunized with either O or HO antigens of either Ogawa or Inaba serotypes contained brucella agglutinins. This activity was absorbed completely from Ogawa antisera by either Ogawa or Inaba O antigens but only partially from Inaba antisera by Ogawa O antigen. These findings support the claim of Gallut that the cross-reaction is due to heat-stable O antigens of
V. cholerae
rather than heat-labile flagellar antigens as described in many text books. The cross-reactive component is more dominant in the Inaba than in the Ogawa serotype of
V. cholerae
.
High Molecular Weight Phosphorus Compound in Nucleic Acid Extracts of the Slime Mold
Physarum polycephalum
Authors: Sauer H. W. , Babcock K. L. , Rusch H. P.
Published: January 2020

Orthophosphate labeled with
32
P was added to the growth medium of the plasmodia of
Physarum polycephalum
. The nucleic acid extracts of such plasmodia contained
32
P that was not removed by nuclease, protease, or amylase. This labeled material was shown to be separable from nucleic acids, could be eluted from a methylated albumin-kieselguhr column at 0.5
m
NaCl, was of high molecular weight, and had several characteristics in common with polyphosphate. A fraction of this polyphosphate-like material was also found in extracts of isolated nuclei.
Hexuronic Acid Dehydrogenase of
Agrobacterium tumefaciens
Authors: Chang Yung Feng , Feingold David Sidney
Published: January 2020

Growth of
Agrobacterium tumefaciens
on
d
-glucuronic acid (GlcUA) or
d
-galacturonic acid (GalUA) induces formation of hexuronic acid dehydrogenase [
d
-aldohexuronic acid: nicotinamide adenine dinucleotide (NAD) oxidoreductase]. The dehydrogenase, which irreversibly converts GlcUA or GalUA to the corresponding hexaric acid with the concomitant reduction of NAD, but not of nicotinamide adenine dinucleotide phosphate was purified 60-fold by MnCl
2
treatment, (NH
4
)
2
SO
4
fractionation, chromatography on diethylaminoethyl Sephadex and negative adsorption with Ca
3
(PO
4
)
2
gel. The
p
H optimum is 8.0. Other uronic acids, aldohexoses, aldopentoses, and polyols, are not substrates. Reduced nicotinamide adenine dinucleotide is an inhibitor strictly competitive with NAD. Kinetic data indicate that the dehydrogenase induced by growth on GlcUA may not be identical with that induced by growth on GalUA.
Differential Lytic Response of Enterococci Associated with Addition Order of Lysozyme and Anions
Authors: Metcalf Robert H. , Deibel R. H.
Published: January 2020

Suspensions ofStreptococcus faecium, prepared by washing with and resuspending in water, were lysed slowly if sodium chloride was added prior to lysozyme; however, if brief incubation with lysozyme was followed by addition of sodium chloride, lysis was immediate and extensive. Relatively lysozyme-resistant strains ofS. faecaliscould be lysed readily by adding lysozyme first. The primary addition of lysozyme apparently resulted in a “sensitized” cell with a damaged wall, as evidenced byN-acetylhexosamine release. Anionic detergents could replace sodium chloride in lysing these sensitized cells. The difference in activity associated with the order of addition probably involved a competition for reactive sites on the cell surface.
Specificity and Efficiency of Thymidine Incorporation in
Escherichia coli
Lacking Thymidine Phosphorylase
Authors: Fangman Walton L.
Published: January 2020

A mutant of
Escherichia coli
lacking the catabolic enzyme thymidine phosphorylase readily incorporates exogenous thymidine into deoxyribonucleic acid (DNA) even when provided at concentrations as low as 0.2 μg/ml. Incorporation by this prototrophic strain occurs specifically into DNA, since, with radioactively labeled thymidine, (i) more than 98% is incorporated into alkali-stable material, (ii) at least 90% is recovered as thymine after brief formic acid hydrolysis, and (iii) at least 90% is incorporated into material with the buoyant density of DNA. During growth in medium containing thymidine, the bacteria obtain approximately half of their DNA thymines from the exogenous thymidine and half from endogenous synthesis. The thymines and cytosines of DNA can be simultaneously and specifically labeled by thymidine-
2
-
14
C
and uridine-
5
-
3
H
, respectively. The mutant, which does not degrade thymidine, retains the ability to degrade the thymidine analogue 5-bromodeoxyuridine.
Regulation of 3-Deoxy-
d
-
arabino
-Heptulosonic 7-Phosphate Acid Synthetase Activity in Relation to the Synthesis of the Aromatic Vitamins in
Escherichia coli
K-12
Authors: Wallace B. J. , Pittard J.
Published: January 2020

Both in vivo and in vitro experiments on wild-type
Escherichia coli
K-12 and mutant strains possessing only single 3-deoxy-
d
-
arabino
-heptulosonic 7-phosphate acid (DAHP) synthetase isoenzymes indicated that, under conditions when all three isoenzymes are fully repressed, sufficient chorismate is still formed for the synthesis of aromatic vitamins. Under repressed conditions both DAHP synthetase (phe) and (trp), but not DAHP synthetase (tyr), were shown to contribute to vitamin production.
Dissimilation of Methionine by a Demethiolase of
Aspergillus
Species
Authors: Ruiz-Herrera José , Starkey Robert L.
Published: January 2020

Enzyme preparations obtained from the mycelium of
Aspergillus
species broke down methionine by co-dissimilation. The deaminase and demethiolase activities of crude extracts were increased 100-fold by precipitation with (NH
4
)
2
SO
4
and column chromatography on diethylaminoethyl cellulose. The enzyme acted on
d
-methionine but not on
l
-methionine. The enzyme was labile: it was inactivated by oxygen and ascorbic acid but ethylenediaminetetraacetic acid and mercaptoethanol preserved its activity. Enzyme activity decreased even at 4 and −30 C and was lost rapidly above 45 C. It was most rapid at 35 C and at
p
H 8.0 to 9.0. For the following reasons, it was concluded that deamination and demethiolation of methionine were effected by the same enzyme: both activities increased equally at each stage of purification; ammonia, methanethiol, and α-keto butyric acid were formed in amounts equivalent to the amount of methionine dissimilated; the
K
m
and optimal
p
H for formation of both keto acid and methanethiol were the same; both activities remained in the same fractions that were separated by electrophoresis and the activities were equivalent. The purified enzyme demethiolated α-keto methionine and α-hydroxy methionine and split the sulfur linkage of ethionine but did not cleave cystathionine. Few amino acids were deaminated. The enzyme was sensitive to some carbonyl and sulfhydryl reagents and was relatively insensitive to heavy metals other than Hg
++
. The
K
m
was 1.3 × 10
−3
to 1.5 × 10
−3
m
at
p
H 7.0. No requirement for cofactors was noted, and attempts to dissociate the enzyme, including dialysis with hydroxylamine, were unsuccessful.
Peptide Synthesis by Extracts from
Bacillus subtilis
Spores
Authors: Bishop Helen L. , Migita Lloyd K. , Doi Roy H.
Published: January 2020

Cell-free peptide synthesis by extracts from vegetative cells and spores of
Bacillus subtilis
was analyzed and compared. The initial rate of phenylalanine incorporation in a polyuridylate-directed system was found to be in a similar range for the two extracts. However, spore extracts frequently incorporated less total phenylalanine as did the vegetative cell system. Optimal conditions for amino acid incorporation by spore extracts were found to be similar to those of vegetative cell extracts. Polyphenylalanine synthesis was stimulated by preincubation of both extracts prior to the addition of polyuridylic acid (poly U) and labeled phenylalanine. Both systems showed a dependence on an energy-generating system and were inhibited by chloramphenicol and puromycin. Ribonuclease, but not deoxyribonuclease, inhibited the reaction significantly. The presence of methionine transfer ribonucleic acid (tRNA
F
) and methionyl-tRNA
F
transformylase was demonstrated in spore extracts. An analysis of several aminoacyl-tRNAs in spores revealed that the relative amounts of these tRNAs were similar to those found in vegetative cells. Only lysine tRNA was found to be present in relatively greater amounts in spores. These results indicate that dormant spores of
B. subtilis
contain the machinery for the translation of genetic information.
Leakiness of Pleiotropic Maltose-negative, Bacteriophage λ-resistant Mutants of
Escherichia coli
K-12
Authors: Ronen A. , Zehavi A.
Published: January 2020

Cultures of
Escherichia coli
K-12
malA
−
λ
r
gal
−
can be transduced to
gal
+
by bacteriophage λ
dg
because of leakiness of the λ
r
phenotype. The efficiency of such transduction is about 10
−5
that of transduction of
mal
+
λ
s
bacteria. Leaky cells (λ
s
phenocopies
) adsorb only very few phage particles, and many transductants, therefore, are defective heterogenotes or show integration of the
gal
+
gene, which is unaccompanied by lysogenization.
Complementation in Nonconidiating Mutants of
Trichoderma
Authors: Weinman-Greenshpan Dvora , Galun Esra
Published: January 2020

Nonconidiating (Con
−
) mutants were isolated from wild-type and color mutants of the fungus
Trichoderma viride
Pers. ex Fries. Heterokaryons were easily produced and maintained, and the complementation relationships among the Con
−
mutants were established. Most Con
−
mutants could complement one or more of the other Con
−
mutants. When marked Con
−
mutants were mixed with marked Con
+
testers, conidiating heterokaryons were formed. The conidia thus obtained produced only the parental type colonies after replating, indicating that nonconidiation is a nuclear characteristic. Allowing two Con
−
colonies to meet and produce a heterokaryon, it was found that the migration of nuclei reached a rate of 5 mm per hr, which is several times greater than the rate of hyphal elongation; it was also found that heterokaryosis of a mycelial region preceded its ability to conidiate.
Ultrastructure of Putrefactive Anaerobe 3679h During Sporulation
Authors: Santo Leatrice Miura , Hohl Hans R. , Frank Hilmer A.
Published: January 2020

Sporulation of putrefactive anaerobe 3679h was studied by observation of ultra-thin sections in the electron microscope. The customary stages were observed during forespore formation, spore maturation, and liberation of the free spore. The exosporium was partially laminated and devoid of the hairlike surface projections observed on spores of some species. Atypical membrane configurations occurred in some cells, and, occasionally, cells with bipolar forespores were found.
Cation-activated Nucleotidase in Cell Envelopes of a Marine Bacterium
Authors: Thompson J. , Green M. L. , Happold F. C.
Published: January 2020

Isolated cell envelopes of a marine bacterium, M.B.3, have been prepared which possess a nonspecific, cation-activated nucleotidase. The cell envelope comprises approximately 35% (dry weight) of the whole cell and contains protein, 60.2%; lipids, 20.7%; hexose, 3.4%; and ribonucleic acid, 4.6%. No deoxyribonucleic acid could be detected in the preparations. The nucleotidase has an essential requirement for Mg
2+
; maximum activation at
p
H 8.0 occurs at a divalent cation concentration of approximately 80 m
m
. At a Mg
2+
to adenosine 5′-triphosphate (ATP) ratio of 2:1, the enzyme was further stimulated by monovalent cations Na
+
, K
+
, NH
4
+
, and Li
+
. Maximum activity was found at a monovalent ion concentration of approximately 0.3
m
. The envelope preparation liberated inorganic orthophosphate (P
i
) from ATP, adenosine 5′-diphosphate (ADP), and adenosine 5′-monophosphate (AMP) at similar rates. Thin-layer and ion-exchange chromatography show that when AMP, ADP, and ATP were utilized as substrate, approximately 1, 2, and 3 moles of P
i
, respectively, were produced per mole of adenosine. P
i
was also liberated from the 5′-triphosphates of guanosine, uridine, and cytidine. The enzyme preparation did not attack
p
-nitrophenyl phosphate, β-glycerophosphate, or inorganic pyrophosphate. Sulfhydryl inhibitors
p
-chloromercuribenzoate,
N
-ethyl maleimide, and iodoacetate had little effect upon the nucleotidase activity. Ca
2+
and ethylenediaminetetraacetic acid caused complete inhibition of the system, whereas ouabain had no effect upon the enzyme activity. The concentrations of Na
+
(0.3
m
) and Mg
2+
ions (60 to 80 m
m
) required for maximum ATP-hydrolyzing activity were similar to those concentrations necessary for maintenance of cell integrity and for the prevention of cell lysis.
Morphological Changes of
Mycobacterium lepraemurium
Grown in Cultures of Mouse Peritoneal Macrophages
Authors: Chang Y. T. , Andersen R. N.
Published: January 2020

Studies were made on morphological changes of
Mycobacterium lepraemurium
grown in cultures of mouse peritoneal macrophages. Two types of nonsolid or irregularly stained
M. lepraemurium
were observed. One type occurred in the growth phase of the organisms during the stage of preparation for bacillary multiplication. The nonsolid bacilli appeared as elongated organisms having pointed ends, isolated acid-fast dots, or faintly stained areas at the ends of the bacilli. It is possible that this irregularity in staining is due to a very gradual, versus an instantaneous, acquisition of acid-fast material during bacillary multiplication and maturation. Solid forms were again observed upon maturation. Nonsolid bacilli were also observed in macrophage cultures infected with autoclave-killed
M. lepraemurium
. Under these conditions there was an emergence of organisms which showed irregularly stained areas and various forms of deformity unaccompanied by elongation or multiplication. These irregularities were most probably due to the destructive process of digestion of bacillary protoplasm. The present study does not support the current hypothesis that all nonsolid acid-fast organisms are nonviable.
Serine Biosynthesis in
Desulfovibrio desulfuricans
Authors: Germano Geno J. , Anderson Kenneth E.
Published: January 2020

Cell-free extracts of
Desulfovibrio desulfuricans
possess enzymes which catalyze the synthesis of serine from 3-phosphoglycerate via the intermediates phosphohydroxypyruvate and phosphoserine.
Evidence for a Reductive Pathway for the Anaerobic Metabolism of Benzoate
Authors: Guyer Mark , Hegeman George
Published: January 2020

Evidence is presented for a reductive pathway for the anaerobic metabolism of benzoate by
Rhodopseudomonas palustris
.
Growth of Parasitic
Mycoplasma
Without Serum or Serum Fraction
Authors: Kurzepa Henryka , Flinton Linda , VanDemark P. J.
Published: January 2020

Lipid-free beef heart infusion would support the growth of six strains of parasitic
Mycoplasma
when supplemented with a combination of cholesterol, lecithin, and cardiolipin.
Unusual
Enterobacteriaceae
: a
Salmonella cubana
that is Urease Positive
Authors: Farmer J. J. , McWhorter Alma C. , Huntley Geraldine A. , Catignani Joan
Published: January 2020

This is the first report of a naturally occurring
Salmonella
that is urea positive. The strain was identified as
Salmonella cubana
and it was typical in all biochemical, serological, and bacteriophage reactions, except that is produced urease strongly.
Evaluation of the Bactrol Disks, a Set of Quality Control Cultures
Authors: Rhoden Dwane L. , Tomfohrde Karla M. , Swenson Jana M. , Smith Peter B. , Balows Albert , Thornsberry Clyde
Published: January 2020

A set of commercially available quality control cultures was evaluated. These cultures were found to be an acceptable alternate to lyophilized or continuously subcultured cultures for a quality control program.
Isolation of infectious bovine rhinotracheitis virus from Mustelidae
Authors: Porter D D , Larsen A E , Cox N A
Published: January 2020

Herpesviruses isolated from either domesticated or wild carnivores should be immunologically compared with known viruses of this group which could have been included in the diet of the animal before being considered to be previously undescribed herpesviruses native to the carnivore in question.
Production of Human Lymphoblastoid Interferon
Authors: Strander H , Mogensen K. E. , Cantell K.
Published: January 2020

The interferon response of 21 lines of human lymphoblasts varied greatly. Interferon from the best producer (11,000 U/ml) resembled human leukocyte interferon.
Recognition of Pseudomonas pickettii in the clinical laboratory: biochemical characterization of 62 strains
Authors: Riley P S , Weaver R E
Published: January 2020

Pseudomonas pickettii strains were studied to determine the characteristics essential for their identification in the clinical microbiology laboratory. Preliminary investigations indicated that these glucose-oxidizing, denitrifying, gram-negative rods were quite similar to an unclassified group of clinical isolates designated VA-2. Gas liquid chromatography of trimethylsilyl derivatives of whole cell hydrolysates of P. pickettii and VA-2 strains yielded nearly identical elution profiles. The VA-2 cultures were concluded to be probable strains of P. pickettii. A method is presented for differentiating cultures of P. pickettii from other similar bacteria encountered in clinical specimens.
Identification of Actinomyces viscosus from canine infections
Authors: Davenport A A , Carter G R , Patterson M J
Published: January 2020

Actinomyces viscosus is a gram-positive, non-acid-fact, facultative, catalase-positive, filamentous, or diphtheroidal microorganism. It was isolated from six canine infections during a period of 1.5 years. The organism was cultured from exudate and flaky granules aspirated from infectious granulomas and empyemas. All cultures grew well aerobically and anaerobically with the addition of 10% carbon dioxide. They fermented lactose, produced catalase and acetylmethylcarbinol, reduced nitrates, hydrolyzed aesculin, and did not produce gelatinase or urease. These physiological characteristics distinguish A. viscosus from other morphologically similar organisms.
Evaluation of blood culture media supplemented with sucrose or with cysteine
Authors: Washington J A , Hall M M , Warren E
Published: January 2020

A total of 5,883 blood samples from patients with suspected bacteremia were inoculated concurrently into each of three media under vacuum with CO2: tryptic soy broth (TSB) with sodium polyanetholesulfonate (SPS), TSB with SPS and cysteine, and TSB with SPS and sucrose. There were 395 positive cultures, excluding presumed contaminants. No significant differences were noted with the addition of cysteine to TSB with SPS, and no streptococcal mutants requiring thiol groups were isolated. Haemophilus, Staphylococcus aureus, and bacteriodaceae were isolated more frequently (P less than 0.05) in the absence of sucrose. The addition of sucrose to TSB containing SPS did not significantly increase the rate of positivity or the time interval to detection of positivity of any group of bacteria.
Simplified scheme for routine identification of human Staphylococcus species
Authors: Kloos W E , Schleifer K H
Published: January 2020

From a total of 40 characters that were previously used to differentiate species of staphylococci, 13 key characters were selected to make a simplified scheme that could be easily used by the routine clinical laboratory for identifying human staphylococci. These key characters included coagulase activity, hemolysis, nitrate reduction, and aerobic acid production from fructose, xylose, arabinose, ribose, maltose, lactose, sucrose, trehalose, mannitol, and xylitol. In the simplified scheme, 924 strains of staphylococci were placed into 11 positions, each of which contained the major portion (greater than or equal to 80%) of strains of one of the recognized species. Several positions contained a rare or few uncommon strains of one or more additional species and these could be resolved on the basis of other key characters.
Purification of Japanese encephalitis virus vaccine by zonal centrifugation
Authors: Okuda K , Ito K , Miyake K , Morita M , Ogonuki M
Published: January 2020

A large-scale procedure for purification and concentration of Japanese encephalitis virus vaccine by a continuous-flow isopycnic banding technique in a sucrose density gradient solution, using a K-III zonal centrifuge rotor, is presented. The quality of zonal-purified vaccine was compared with commercial and Japanese National Institutes of Health reference vaccines for antigenicity, immunodiffusion, and allergic encephalitogenicity tests to show its high purity.
Comparison of recovery rates of various organisms from clinical hypertonic blood cultures by using various concentrations of sodium polyanethol sulfonate
Authors: Rosner R
Published: January 2020

By using parallel culture techniques, the recovery rates of a wide spectrum of organisms encountered in hypertonic clinical blood cultures was determined from four different blood culture bottles. Each bottle was identical except for the amount of sodium polyanethol sulfonate (SPS) present. Flasks A, B, C, and D contained SPS in final concentrations of 0.025, 0.05, 0.075, and 0%, respectively. Of 144 patients found to have clinically relevant organisms in their blood cultures, 127 had positive A flasks, 144 had positive B flasks, 140 had positive C flasks, and 110 had positive D flasks. There was no significant difference in the time required to obtain organism recovery from the A, B, or C flasks; however, the time required to obtain organism recovery from the D flask was considerably longer, ranging up to 5 days in many cases. Of the various organisms recovered, 3 of 7 strains of anaerobic streptococci and 1 of 28 strains of Streptococcus pneumoniae appeared to be inhibited by SPS when the concentration was 0.075%. In no case was an organism recovered from either the A or D flask but not from the B flask, indicating that a concentration of 0.05% SPS in hypertonic media does not inhibit the growth of a wide spectrum of organisms in clinical blood cultures.
Effect of acetone fixation on infectivity and antigenicity of respiratory syncytial virus and adenovirus in the fluorescent antibody test
Authors: Bardell D
Published: January 2020

An investigation was made on the effect of acetone fixation on infectivity and immunofluorescent antigenicity of respiratory syncytial virus and adenovirus type 5, lipid- and nonlipid-containing viruses, respectively. Viruses were allowed to replicate in HEp-2 cells, and the cells were then fixed in acetone at 5 C for periods ranging from 30 s to 7 days. Treatment for 10 min was sufficient to inactivate respiratory syncytial virus, whereas infectious adenovirus type 5 could be isolated from cells immersed in acetone for 7 days. There was a gradual reduction, to 50% of that observed at 30 s, in the intensity of fluorescent antibody staining of both viruses with increasing fixation time, but no significant decreases in fluorescent antibody end-point titers of antisera to either virus were observed.
Improved Epstein-Barr virus immunoglobulin M antibody test
Authors: Joncas J H , Granger-Julien M , Gervais F
Published: January 2020

The successful demonstration of Epstein-Barr virus immunoglobulin M (EBV IgM) antibody in human sera has been accomplished to date by at least four groups of workers. Many, however, including ourselves, have had difficulty in getting reproducible results with the techniques described. The three-coat technique described by H. Schmitz and M. Scherer (1972) on both fractionated and unfractioned sera was adopted with minor modifications. The Hyland antihuman IgM antiserum used in the second coat was made specific by absorption on Cohn fraction II. This step in the procedure was found to be the single most important factor in arriving at reproducible results in the IgM test. The EBV IgM antibodies from our results to date with this method in 14 cases of heterophil-positive cases of mononucleosis appear short lived, lasting 2 months or less. These antibodies were found in only 2 of 18 selected non-mononucleosis cases, in both associated with EBV-viral capsid antigen antibody rise or seroconversion. The successful elimination of nonspecific fluorescence by a simple, inexpensive procedure and the possibility of testing unabsorbed, unfractionated sera directly will facilitate the use oe the EBV IgM antibody test in the future.
Methods for the detection of viruses in bovine serum
Authors: Swack N S , Fong C K , Hsiung G D , Gross P A
Published: January 2020

An evaluation of selected commonly used procedures for the recovery of endogenous viral contaminants in bovine serum was undertaken. Low speen centrifugation (25,000 x g) was found to be efficient for the recovery of bovine herpesvirus type 1 (BHV-1) and parainfluenza virus type 3(PI-3) in bovine serum. Decreased infectivity titers were obtained when parainfluenza virus type 3, and to a lesser extent bovine herpes virus type 1, were concentrated using high speed centrifugation (100,000 x g) for extended time periods. In neither case could infectious virus be recovered from serum containing sufficient titers of homologous neutralizing antibody, although electron microscopy examination revealed the presence of the viruses previously added. In the presence of homologous antibody, virus particles appeared to have a diffuse, poorly defined outer membrane. Neutralizing antibody titers to bovine herpesvirus type 1 and outer membrane. Neutralizing antibody titers to bovine herpesvirus type 1 and parainfluenza virus types were found in fetal, calf, and adult bovine sera. The prevalence and magnitude of the antibody titers to these viruses increased with the age of the animals examined.
Significant Enhancement of T-Strain Mycoplasmal Growth on Supplemented Shepard's A6 Differential Agar
Authors: Tull A. H. , Blair E. B. , Fishman D. L. , Heatley G. J.
Published: January 2020

Growth of T-strain mycoplasmas was significantly improved by supplementing Shepard's A6 medium with sodium thioglycolate, cysteine hydrochloride, mercaptoethanol, or dithiothreitol.
Plaque Assay for Rabies Serogroup Viruses in Vero Cells
Authors: Buckley Sonja M. , Tignor G. H.
Published: January 2020

Plaque formation in Vero cells has been induced with seven rabies serogroup viruses either cocultivated as infected BHK-21 or
Aedes albopictus
cells, or directly cultivated as infected mouse brain.
Sensitivity of bhk-21 cells supplemented with diethylaminoethyl-dextran for detection of street rabies virus in saliva samples
Authors: Larghi O P , Nebel A E , Lazaro L , Savy V L
Published: January 2020

A tissue culture system for detecting rabies virus from saliva samples of suspected animals was developed and compared to suckling mouse inoculation. Swab samples were obtained from the mouth of the animal heads received for rabies diagnosis; these swabs were submerged in maintenance medium. The maintenance medium was inoculated intracerebrally into suckling mice and onto BHK-21 cells with diethylaminoethyl (DEAE)-dextran (BHK/DEAE) and without (BHK). Rabies immunofluorescence was performed on the brain of the mice dying during the observation period and also on both tissue culture systems every day after infection. The BHK-DEAE system detected 28 positive samples obtained from 48 rabid animals and the BHK system detected 18. By suckling mouse inoculation only 11 of the same positive samples were detected. A total of 90 samples was studied by the three methods. Rabies virus was detected by the tissue culture methods earlier than by suckling mouse inoculation. The BHK-DEAE method was an economic and fast method for rabies virus detection in saliva samples, which could be used for ecological and pathogenesis studies, as well for rabies diagnosis before the death of the suspected animal.
Suitability of peracetic acid for sterilization of media for mycoplasma cultures
Authors: Wutzler P , Sprössig M , Peterseim H
Published: January 2020

The utility of peracetic acid for sterilization of serum and yeast extract additions to mycoplasma medium was studied by culturing six Mycoplasma species. Culture media containing additions that had been sterilized with peracetic acid proved to be as good as filtered components. The use of 0.05 to 0.1% peracetic acid is recommended to sterilize the serum and yeast extract additions since savings in time and equipment can be accomplished.
Simplified microimmunofluorescence test with trachoma-lymphogranuloma venereum (Chlamydia trachomatis) antigens for use as a screening test for antibody
Authors: Wang S P , Grayston J T , Alexander E R , Holmes K K
Published: January 2020

A simplified microimmunofluorescence test with trachoma-lymphogranuloma venereum (Chlamydia trachomatis) antigens has been devised as a screening test for antibody in human sera. The test differs from our standard procedure by amalgamating 15 different immunotypes into nine antigen pools, by using only three serum dilutions, and by dropping use of duplicate slides. The screening test could be performed on at least six times as many sera as the standard test for a given unit of effort. It was shown to have a sensitivity equal to the standard test and an ability to determine specific immunotype on the basis of antibody pattern (72%) only slightly reduced from the standard test (84%). The screening test was carried out on 876 patients from different population groups in the Seattle area. The tests showed that antibody was present frequently in persons attending venereal disease clinics (60%) and commonly (25%) in a group of adults without venereal disease. Nine percent of children under 15 years of age tested had antibody; the significance of this finding is unknown.
Virulence factors of biotypes of Staphylococcus epidermidis from clinical sources
Authors: Males B M , Rogers W A , Parisi J T
Published: January 2020

The biotyping scheme of Baird-Parker was applied to cultures of Staphylococcus epidermidis from patients. In all, 63.6% of 228 cultures belonged to biotype 1, followed by biotypes 4, 3, and 2 in decreasing order of incidence. When classified according to clinical source of isolation, cultures of S. epidermidis were most frequently isolated from urine, with 39.5% of 228 cultures from this source. Each of the four biotypes was distributed throughout all nine catagories of clinical sources. The production of virulence factors was based on the results of three groups of tests: (i) deoxyribonuclease, urease, gelatinase, caseinase, and lysozyme production; (ii) lipolytic activity on the tweens; and (iii) hemolysin production. Enzymatic activity was highest for organisms in biotypes 1, followed by biotypes 3, 4, and 2 in decreasing order. Of the 228 cultures, 76.3% were lysed by lysostaphin. Resistance to antibiotics was highest for tetracycline, ampicillin, and penicillin, with rates of 54.8, 69.3, and 81.6%, respectively. The role of S. epidermidis as an etiological agent was studied by analyzing the laboratory and clinical data of 80 patients selected at random with bacteriuric S. epidermidis. Organisms in biotype 1 were most commonly associated with urinary tract infection. The significance of certain biotypes of S. epidermidis as opportunistic pathogens among compromised hosts in a hospital environment is discussed.
Complement-fixing antibody in human sera reactive with viral and soluble antigens of cytomegalovirus
Authors: Cremer N E , Schmidt N J , Jensen F , Hoffman M , Oshiro L S , Lennette E H
Published: January 2020

Antibody titers to cytomegalovirus were determined in 204 single-serum specimens and in 138 serum pairs, utilizing as antigen extracts of infected cells prepared by either freezing and thawing cells (FT antigen) or extracting them with a glycine buffer (GE antigen). With the serum pairs, 40% of the early sera and 44% of the late sera showed a fourfold or higher antibody titer with the GE antigen compared with those with the FT antigen, whereas 39% of the single sera had a fourfold or higher titer with the GE antigen. Sucrose density gradient centrifugation of the two antigen preparations indicated that the GE antigen contained two peaks of complement-fixing activity, one at a density of 1.18 to 1.22 g/cm3 and one at 1.06 to 1.10 g/cm3. The FT antigen had only one discernible peak of complement-fixing activity at 1.04 to 1.10 g/cm3. On electron microscopy, the more dense peak contained mainly nucleocapsids with some dense bodies and occasional enveloped virions, and the lighter peak was composed of amorphous material. Antibody titers obtained with the crude GE antigen and the more dense peak (designated viral antigen) were in agreement, whereas titers with the FT antigen agreed with those obtained with the lighter peak (soluble antigen). The antibody detected with both antigens was mainly immunoglobulin G.
Passive transfer of immunity into leprosy patients by transfusion of lymphocytes and by transfusion of Lawrence's transfer factor
Authors: Saha K , Mittal M M , Maheshwari H B
Published: January 2020

About 1,200 million viable lymphocytes from normal but lepromin- and tuberculin-positive human beings were transfused in four patients of lepromatous and one of tuberculoid leprosy three times at monthly intervals. Three patients of lepromatous leprosy suffered from erythema modosum, whereas the other two developed severe reaction whenever put on the smallest dose of dapsone. In one patient of lepromatous leprosy, minimal improvement or none was observed, whereas in the remaining three cases of lepromatous and one of tuberculoid leprosy, clinical, bacteriological, as well as histological improvement occurred. Two of the five patients started to tolerate the dapsone during the period of study. The present study indicates that immunotherapy might have a definite role in the management of the disease especially in cases with erythema nodosum. Lawrence factor, prepared from leucocytes of normal donors, was transfused three times into four lepromatous leprosy patients who were intolerant to anti-leprosy drugs. The donors were healthy but were tuberculin and lepromin (Mitsuda) positive. The clinical, histological, bacteriological (morphological index), and immunological assessments of the patients were performed before and 5 months after starting the immunotherapy. In two patients conversion of Mitsuda reaction occurred, but there was no appreciable improvement in the clinical, histologic, and bacteriologic status of these patients.
Detection by radioimmunoassay of antibodies in human smallpox patients and vaccinees
Authors: Ziegler D W , Hutchinson H D , Koplan J P , Nakano J H
Published: January 2020

A radioimmunoassay procedure was developed for determining smallpox and vaccinia antibodies in human sera. The test detected and measured both primary and secondary immune responses in persons infected with variola virus or vaccinia virus. The antibody titers obtained by complement fixation, hemagglutination inhibition, plaque reduction neutralization, and radioimmunoassay methods were compared. In sequential serum specimens, the radioimmunoassay test indicated fourfold or greater increases in all of the smallpox patients and in six of eight vaccinated persons. Both the complement fixation and the hemagglutination inhibition tests were less effective. In persons who had been vaccinated, radioimmunoassay and plaque reduction neutralization tests appeared to measure the same immune response. However, in smallpox patients the immune response was readily detected by radioimmunoassay, whereas an immune response was not detected by the plaque reduction neutralization test when vaccinia virus was the antigen in the test system. Radioimmunoassay is an operationally simple procedure which provides objective and quantitative end-point titers in serological determinations.
Diagnosis of Herpesvirus hominis infections in a general hospital laboratory
Authors: Hanna L , Keshishyan H , Jawetz E , Coleman V R
Published: January 2020

There is a greatly increased interest in Herpesvirus hominis infections especially those of type 2 associated with genital lesions or neonatal disease. Physicians are eager to confirm the clinical impression with a specific virologic diagnosis such as isolation of the agent and its typing, and type-specific antibody responses. Procedures are reviewed here which permit such studies in general microbiology laboratories equipped for simple cell culture and immunofluorescence. This paper recounts experience with several laboratory methods and evaluates their efficiency and practicability in a general laboratory. Virus isolation was optimal if specimens were obtained from visible lesions early in their evolution and it often provided a specific diagnosis, including typing of the isolate by immunofluorescence, within 24 to 48 h. Estimation of serum antibodies to herpes simplex virus type 1 and 2 by indirect immunofluorescence was more sensitive and perhaps also more specific than by microneutralization test. A pilot study of herpes simplex virus antibody titers in mothers and in the cord blood of the offspring suggested the need to evaluate a possible protective role of high titer antibody in the fetus.
Cultural and biochemical characteristics and fatty acid composition of Pseudomonas diminuta and Pseudomonas vesiculare
Authors: Kaltenbach C M , Moss C W , Weaver R E
Published: January 2020

The cultural characteristics, biochemical activity, and cellular fatty acid composition of Pseudomonas diminuta and Pseudomonas vesiculare provided means for differentiation of these closely related species. Broth cultures of P. diminuta showed turbid growth and a distinct surface pellicle after 24 h at 35 C. P. vesiculare had no pellicle, and only light, diffuse growth was observed. All strains of P. vesiculare oxidized maltose and hydrolyzed esculin to varying degrees; P. diminuta was negative in these tests. The cellular fatty acids of these two species were similar, except that P. diminuta possessed a C19 cyclopropane acid which was not detected in P. vesiculare.
Screen for type-C ribonucleic acid viruses in vaccines using the ribonucleic acid-dependent deoxyribonucleic acid polymerase assay
Authors: Milstien J B , Petricciani J C
Published: January 2020

The ribonucleic acid-dependent deoxyribonucleic acid polymerase assay was used to detect type-C viruses in live virus vaccines. Conditions were first established to maximize the sensitivity of the assay. Vaccines tested included live poliomyelitis, rubella, measles, mumps, and yellow fever. Only yellow fever and measles vaccines known to have been produced in avian leukosis-contaminated cells showed evidence of type-C viruses using the assay. The result of the survey show that the assay has direct practical application to the problem of detecting latent agents in biological products intended for human use.
New human adenovirus isolated from a renal transplant recipient: description and characterization of candiate adenovirus type 34
Authors: Hierholzer J C , Atuk N O , Gwaltney J M
Published: January 2020

An antigenically distinct adenovirus is described which was isolated in March 1972 from the urine of a 17-year-old Caucasian male who was experiencing fever after receiving a kidney transplant from a cadaver in February. The adenovirus could not be isolated in April from a pharyngeal swab which yielded cytomegalovirus. Complement-fixation, hemagglutination-inhibition, and/or serum-neutralization tests on sequential serum specimens from the patient confirmed that the adenovirus infection occurred during March and showed that infections with cytomegalovirus and respiratory syncytial virus also occurred during late March and April. The patient's persistent fever, for which other causes could not be found, may have been associated with one or more of these infections. Upper respiratory symptoms and lung involvement were not found during this period. Mild liver dysfunction during this time could not be clearly related to adenovirus infection because of the presence of multiple other causes. The adenovirus may have been latent in the donor kidney and become active in the new host as a consequence of immunological impairment. The adenovirus, purified by terminal dilution and plaque procedures, has antigenic, morphological, biophysical, host susceptibility, and hemagglutinating properties characteristic of adenovirus group IA. Buoyant densities in CsCl are 1.340 g/ml for the virion, 1.304 g/ml for the group CF antigen (hexon), 1.295 g/ml for the major soluble complete hemagglutinin (dodecon), and 1.206 g/ml for the minor soluble complete hemagglutinin (tentatively, fiber dimer). The virus does not cross-react in reciprocal hemagglutination-inhibition and serum-neutralization tests with antisera to adenovirus types 1 to 33. We propose this virus as candidate adenovirus type 34 (Compton).
Characteristics of Pasteurella multocida of human origin
Authors: Heddleston K L , Wessman G
Published: January 2020

Physiological, serological, morphological, and cultural differences were observed among 30 Pasteurella multocida cultures of human origin. The usual variations in the fermentation of glycerol, lactose, sorbitol, trehalose, and xylose were observed. Unlike most P. multocida, two cultures did not produce indol. Six serotypes were found. In addition to the widely recognized iridescent, blue, and watery mucoid (circular) colonies, punctiform colonies were observed. None of the cultures were pathogenic for turkeys. Results of the study indicate that one should be aware of the many variable characteristicx of P. multocida of human origin to facilitate indentification.