bluuebunny/crossref_metadata_2025 huggingface dataset:
Practical aerobic membrane filtration blood culture technique: development of procedure
Authors: Sullivan N M , Sutter V L , Finegold S M
Published: January 2020

The advantages of a membrane filter system for blood culturing have been realized for many years. Lysing of the blood prior to filtration is a convenient way to proceed, but previously described lysing procedures result in loss of certain organisms, particularly gram-negative bacilli. Four concentrations of Triton X-100 and sodium carbonate were studied in vitro, and their lysing and antibacterial properties were observed. A solution of 0.08% Na2CO3 and 0.005% Triton X-100 was found to have the least antibacterial effect and gave consistently good lysis and filtration times (under 3 min). An 8.3-ml amount of blood added to 190 ml of this concentration of lysing solution, filtered through three 47-mm membrane filters (0.45-mum pore size), led to recovery of 85% or more of various aerobic and facultative organisms in studies of artificially seeded blood.
Practical aerobic membrane filtration blood culture technique: clinical blood culture trial
Authors: Sullivan N M , Sutter V L , Finegold S M
Published: January 2020

During the course of preliminary clinical trials of an improved membrane filter blood culture system, filter plugging produced by a gelling of the lysing solution was observed when the patients had high leukocyte counts. A solution of streptokinase-streptodornase (Varidase, Lederle Laboratories) dissolved or prevented the gel and permitted rapid filtration without plugging. With streptokinase-streptodornase incorporated in the filtration procedure, a comparison of several culture systems was carried out on 176 blood cultures. Brucella broth with and without sodium polyanethol sulfonate, a prereduced osmotically stabilized broth, pour plates, and an improved aerobic membrane filter system were compared. The membrane filter system yielded 29 of the total of 37 positive cultures, far surpassing all other systems. Eight of these cultures were detected first by the filter technique, and 13 were positive only in this system. Nineteen of the 37 positive cultures were from patients on antimicrobial agents. Fourteen of these were detected by the filter, twice the number detected by any of the other systems.
Simple amidase test for identification of mycobacteria
Authors: Helbecque D M , Handzel V , Eidus L
Published: January 2020

A modified amidase test for differentiation of mycobacteria is described. A total of 224 atypical mycobacteria, 154 Mycobacterium tuberculosis, and 26 M. bovis strains were classified by this procedure. Of the 404 strains of various species studied, 400 exhibited an amidase spectrum identical to the established pattern. The simplicity of this method may promote its application in routine examinations.
Transmission of Lactobacillus jensenii and Lactobacillus acidophilus from mother to child at time of delivery
Authors: Carlsson J , Gothefors L
Published: January 2020

The presence of Lactobacillus jensenii and Lactobacillus acidophilus has been studied in specimens from the rectum and vagina of the mother, from the mouth of the infant at the time of delivery, and from the mouth and rectum of infants six days of age. L. jensenii could be differentiated from other species of lactobacilli by the following combination of characteristics: production of only D-lactate, hydrolysis of arginine, and fermentation of cellobiose, galactose, and ribose, but not of lactose. L. jensenii and L. acidophilus were common inhabitants of the vagina. In spite of a contamination of the infant's mouth by L. jensenii and L. acidophilus during delivery, neither of these organisms became established in the mouth of the newborn infants.
Rapid method to detect rubella immunoglobulin M and immunoglobulin A antibodies
Authors: Schmitz H , Shimizu H , Kampa D , Doerr H W
Published: January 2020

Immunoglobulin (Ig) G was removed from serum specimens by precipitation with gamma chain-specific anti-human IgG of rabbit origin. The remaining rubella virus-specific IgM (and IgA) antibodies were then detected by the rubella hemagglutination-inhibition test. This procedure has proven to be as reliable as estimations carried out with IgM fractions separated on a sucrose density gradient.
Immunofluorescent antibody test for diagnosis of gonorrhoea
Authors: Caloenescu M , Clecner B , Petrow S , Kasatiya S S
Published: January 2020

An indirect fluorescent antibody test was evaluated in 198 cases of a high-risk group with a culture prevalence of 37.3% and in 426 cases of a low-risk group with a culture prevalence of 1.16%. A sensitivity of 77.1% in the culture-positive patients with uncomplicated gonorrhoea, and a specificity of 88.7% in the culture- and history-negative cases, was obtained in the high-risk group. In this group, the sera from 88.8% of the patients with culture-proven gonorrhoea became positive in an indirect fluorescent antibody test within 3 weeks of last sexual contact. In the low-risk group, for which the sensitivity could not be determined due to various reasons, a specificity of 95.8% was obtained. Complement fixation test was positive in sera of only 17.6% of the culture-positive cases of the high-risk group.
Quantitative assay of dermatophyte-infected guinea pig skin scales
Authors: Sinski J T , Pokrifchak J S , Kelley L M
Published: January 2020

A technique to obtain homogeneous suspensions of fungal particles in scrapings from infected skin was devised that permits delivering uniform inocula to culture media being evaluated for suitability as primary isolation media for dermatophytes. The technique consisted of trypsinization of the scales, utilizing scrapings from lesions experimentally induced with an isolate of Trichophyton mentagrophytes var. granulare on guinea pigs. Optimal conditions for dislodging the fungal particles were assessed by colony counts. One-hour treatments with 0.5% (1:300) trypsin yielded the highest counts.
Experiences and observations with the typing of Staphylococcus aureus phage 94
Authors: Marraro R V , Mitchell J L
Published: January 2020

During a 28-month period, 5,078 cultures from a variety of anatomical sites were received for staphylococcal phage typing. Of these, 503 (10%) were not suitable for the procedure requested. Of the 4,575 viable cultures, 1,030 (23%) of the microorganisms were nontypable at both the routine test dilution (RTD) and at 100 times RTD. Of the 3,545 typable organisms, 3,061 (86%) were lysed at RTD, whereas 484 (14%) were typed only at 100 times RTD. Observations pertaining to the typing efficacy of staphylococcal phage 94 indicate that 651 (18%) of the typable microorganisms were lysed only by phage 94 at RTD or at 100 times RTD. Without the addition of this new phage to the international basic set, the number of nontypable strains (1,030 or 23%) would have been 1,681 (39%). Data regarding the geographic distribution of Staphylococcus aureus phage 94 point to the occurrence of the host strain in 13 (68%) of 19 states and 18 (62%) of the 29 hospitals submitting specimens to this laboratory. The assumed origin and speculated mode of dissemination of this microorganism are discussed.
Survival of anaerobic and aerobic bacteria on cotton swabs in three transport systems
Authors: Yrios J W , Balish E , Helstad A , Field C , Inhorn S
Published: January 2020

The capacity of aerobic and anaerobic bacteria to survive on cotton swabs placed into a dry gassed-out CO2-filled tube (DGT), dry sterile aerobic tube (DAT), and a tube containing a modified Stuarts' transport medium (MST), was assessed. Pseudomonas aeruginosa increased in numbers by 2 and 3 logs when stored in MST and DAT, respectively. The viability of P. aeruginosa, although retarded when compared to MSA and DAT, was not adversely affected by the CO2 environment in the DGT. The MST maintained relatively constant numbers of Streptococcus pyogenes during the 48-h storage period. The DAT and the DGT were unable to maintain the viability of S. pyogenes. Staphylococcus aureus, when stored in a DGT, DAT, or MST, was maintained in relatively constant numbers throughout the entire storage period. Of the four anaerobic bacteria evaluated (Bacteroides fragilis ssp. thetaiotaomicron, Bacteroides melaninogenicus ssp. asaccharolyticus, Fusobacterium nucleatum, and Peptostreptococcus anaerobius), only B. fragilis ssp. thetaiotamicron survived the 48-h storage period in the DGT. Under these test conditions the DGT did not adequately maintain the viability of the majority of anaerobic bacteria tested (when held on cotton swabs). However, the MST did maintain the viability of all species tested for at least the first 2 h of storage.
Large-scale production of rubella precipitinogens and their use in the diagnostic laboratory
Authors: Cappel R , Schluederberg A , Horstmann D M
Published: January 2020

A method for producing large quantities of rubella theta and iota precipitating antigens in Vero cells is described. Using this reagent, the detection of anti-theta antibodies was found to be as sensitive as the hemagglutination inhibition test for both the determination of immune status and the diagnosis of acute rubella infection. The detection of anti-iota antibodies may permit diagnosis of rubella infection, when the collection of the first serum has been delayed and the early rise of hemagglutin-inhibiting and anti-theta antibodies has been missed.
Increased selectivity of Mitis-Salivarius agar containing polymyxin
Authors: Fitzgerald R F , Adams B O
Published: January 2020

Incorporation of the antibiotic polymixin B sulfate in plates of Mitis-Salivarius medium at a level of 100 U per ml effectively suppressed the growth of gram-negative spreading organisms which occasionally overgrow Mitis-Salivarius plates of oral samples and make them unsuitable for enumeration and differentiation of streptococci. At the indicated level the antibiotic did not affect the growth or typical colonial characteristics of the oral streptococci.
Inhibition of growth of Coccidioides immitis on Sabouraud medium containing polymyxin B
Authors: Collins M S
Published: January 2020

No growth of Coccidioides immitis occurred when fluid from infected tissue or arthrospores suspended in distilled water were plated on the surface of Sabouraud medium, solidified with refined agar, and containing 20 mg of polymyxin B per liter. Solidification of Sabouraud medium with unrefined agar completely abolished the anticoccidioidal activity of polymyxin B.
Evaluation of the pathotec Rapid I-D system for identification of Enterobacteriaceae
Authors: Smith P B , Rhoden D L , Tomfohrde K M
Published: January 2020

The PathoTec Rapid I-D System for identifying Enterobacteriaceae was evaluated with 471 cultures. In 4,910 individual test comparisons, 95.5% of the results agreed, with results of only two test strips, those for esculin hydrolysis and urease production, agreeing with conventional tests in less than 94% of the trials. The PathoTec system exhibited 94.3% accuracy in identifying these cultures in a double-blind study with conventional media and procedures as the alternate system. Two newly developed test strips, for 0-nitrophenyl-beta-D-galactopyranoside and ornithine decarboxylase, were found to be highly reliable.
Differentiation of Known Strains of BCG from Isolates of
Mycobacterium bovis
and
Mycobacterium tuberculosis
by Using Mycobacteriophage 33D
Authors: Jones Wilbur D.
Published: January 2020

The use of mycobacteriophage 33D to differentiate strains of BCG from isolates of
Mycobacterium bovis
was investigated. The procedure was found to be reproducible and, using the commercially available media described, can be recommended for use in mycobacterial reference laboratories.
Recovery of
Cryptococcus neoformans
from Modified Dubos Liquid Medium Utilized for Isolation of Mycobacteria
Authors: Damsker Beca , Bottone Edward J.
Published: January 2020

Cryptococcus neoformans
was isolated from nine pathological specimens cultured for mycobacteria. Five of these were recovered only in liquid TBC medium in the absence of growth on conventional substrates.
Two serotypes of exfoliatin and their distribution in staphylococcal strains isolated from patients with scalded skin syndrome
Authors: Kondo I , Sakurai S , Sarai Y , Futaki S
Published: January 2020

Two serotypes of exfoliatin, A and B, previously reported by Kondo et al. (1974) were examined for their presence in 43 strains of Staphylococcus aureus, most of which were isolated from patients with Ritter's disease and impetigo. The tested strains consisted of 24 strains of phage group II and 19 strains not of phage group II. Twenty-four strains were found to produce an exfoliatin of either type A or type B, but 16 strains produced both types and 3 strains produced neither. No relationship was found between the serotype of an exfoliatin and the phage type of the exfoliative strain, although the single producers of exfoliatin A were all found to belong to phage group II and those of exfoliatin B to the other phage group.
Comparison of hemoglobin-free culture media and thayer-martin medium for the primary isolation of Neisseria gonorrhoeae
Authors: Jacobs N F , Kraus S J
Published: January 2020

Translucent culture media are essential for the determination of gonococcal colony types. In this study two hemoglobin-free, translucent culture media were compared with Thayer-Martin (TM) medium for the primary isolation of Neisseria gonorrhoeae from male urethral specimens. The antibiotic-free translucent medium (GCB) and the translucent medium containing vancomycin, colistin, and nystatin (VCN) were as efficient as TM medium in detecting positive specimens. In fact, VCN medium gave positive results significantly more often than TM medium(P less than 0.05). The amount of growth on positive plates was comparable for all three media. Contaminants were noted most often on GCB medium (P less than 0.0001), and the frequency of contaminants with TM plates exceeded that of VCN plates (P less than 0.0001). The high isolation rate of N. gonorrhoeae with VCN hemoglobin-free culture medium and its lower cost and greater ease of preparation make it a suitable alternative to TM medium for use with male urethral specimens.
Viral studies on human brain culture retrieved from cold storage
Authors: Lennette E T
Published: January 2020

Tissue cultures established from brain tissues of six patients with various chronic, degenerative diseases did not show evidence of viral agents. Sucrose gradient analysis of radioactively labeled culture fluid from two patients failed to reveal the presence of any incomplete or defective virus. Extracellular fluids from three brain cultures contained a factor able to stimulate deoxyribonucleic acid synthesis of Vero cells.
Serological diagnosis of California (La Crosse) encephalitis by immunofluorescence
Authors: Kalis J M , Burgess A C , Balfour H H
Published: January 2020

A method of indirect immunofluorescence was developed and examined retrospectively as a serological test for the laboratory diagnosis of California encephalitis (CE). LaCrosse virus immunofluorescence immunoglobulin (Ig) G and IgM studies were done on paired sera from 50 patients with acute central nervous system infections. CE had been documented in 25 patients by hemagglutination inhibition, neutralizing, complement fixing, and/or precipitin tests. Five (20%) of the acute and 16 (64%) of the convalescent sera from CE patients had La Crosse IgM antibodies. Seven (28%) of the acute and all of the convalescent CE specimens had La Crosse IgG antibodies. Titers ranged from less than 4 to 256. IgG antibodies were present in all 11 sera collected 1 to 2 years after CE, but IgM antibodies were absent. The 25 serum pairs from patients who did not have CE were negative for IgM and IgG antibodies. This study indicated that La Crosse immunofluorescence antibody tests were as sensitive and specific for CE as conventional hemagglutination-inhibition tests, and would detect at least 20% of patients during their acute illness.
Selective interaction of Neisseria gonorrhoeae and Candida albicans and its possible role in clinical specimens
Authors: Hipp S S , Lawton W D , Savage M , Gaafar H A
Published: January 2020

A study of 27 clinical specimens from which Neisseria gonorrhoeae and Candida albicans were isolated simultaneously indicated that 44% of the gonococcal isolates were resistant to inhibition by the C. albicans with which they were found and an additional 33% were totally resistant to inhibition by all C. albicans tested. All 27 C. albicans showed inhibitory activity against standard indicator strains of N. gonorrhoeae.
Rapid Identification of Swarming
Proteus
Using the PathoTec Ornithine Decarboxylase Test Strip
Authors: Lund Marlys E. , Matsen John M. , Blazevic Donna J.
Published: January 2020

The PathoTec ornithine decarboxylase test strip was evaluated for use in identification of swarming
Proteus.
All 181 strains tested were accurately identified within 4 h.
Hemolysis-in-gel test for the demonstration of antibodies to rubella virus
Authors: Strannegård O , Grillner L , Lindberg I M
Published: January 2020

A rapid and simple method for the determination of rubella immunity is described. The method, which employs passive hemolysis in agarose gel, is sensitive and reproducible and does not require prior absorption of test sera to remove inhibitors. Immunoglobulin G, but not immunoglobulin M, antibodies were regularly found to give demonstrable reactions in the test. It is concluded that the hemolysis-in-gel test may provide a valuable tool, in particular for mass screening for rubella immunity.
Stability of Respiratory Syncytial Virus in a New Bentonite Holding Medium
Authors: Klein Joel D. , Strangert Katherine , Collier Albert M.
Published: January 2020

Stability of respiratory syncytial virus was determined in a new bentonite holding medium and compared with two other commonly used media. Eventual infectivity titers were similar with all three preparations.
Preparation of reference antisera for laboratory diagnosis of blastomycosis
Authors: Green J H , Harrell W K , Johnson J E , Benson R
Published: January 2020

Antiserum was prepared in rabbits against the A antigen and a new antigen (labeled the D antigen) of yeast-phase cells of Blastomyces dermatitidis by using immunoelectrophoretic precipitin arcs as vaccine. The antiserum can be used as a reference serum in the immunodiffusion test for identyfying antibodies to the A antigen in sera from patients thought to have blastomycosis. Antibodies to the D antigen have not yet been found in human sera. The rabbit antiserum that was absorbed with Histoplasma capsulatum yeast-phase cells did not react in the complement fixation test with either H. capsulatum yeast-phase or mycelial-phase complement fixation antigen. Therefore, it could also be used as a positive control serum in the complement fixation test. A fluorescent-antibody conjugate prepared from the absorbed serum was found to be valuable for the identification of yeast-phase cells of B. dermatitidis.
New serotypes of group B streptococci isolated from human sources
Authors: Perch B , Kjems E , Henrichsen
Published: January 2020

Antisera raised in rabbits against two nontypable group B streptococci, which were not agglutinable in a specific group B antiserum, were tested with acid extracts of 78 nontypable human group B streptococci. One antiserum (12351) reacted with 15 strains, and the other (7271) reacted with only 2 strains. Antiserum to Wilkinson's strain SS 1169 (NT 1) reacted with three strains. Antiserum against strain 12351 appears to be a useful antiserum against a new type antigen, which is probably polysaccharide in nature.
API and Minitek systems in identification of clinical isolates of anaerobic gram-negative bacilli and Clostridium species
Authors: Hanson C W , Cassorla R , Martin W J
Published: January 2020

A comparison of the API and Minitek methods of biochemical testing was made on a variety of anaerobic bacteria. Although API and Minitek results were not compared to more standardized or conventional procedures of identification, multiple repeat testing of the two systems was done on routine clinical isolates and known organisms to determine (i) whether the reactions were reliably consistent, (ii) the ease of reading the two systems with respect to the frequency of questionable results, and (iii) the percentage of routine clinical isolates for which each system yielded an identification. The Minitek system gave a much lower incidence of difficult to interpret reactions. The two systems were comparable in terms of reproducibility and capability of yielding an identification of the anaerobic gram-negative bacilli and Clostridium species, but were unsatisfactory for routine use on most of the other anaerobic bacteria isolated.
Variable Neutralization of Several Nonspecific Antibacterial Systems in Fresh, Defibrinated Human Blood by Sodium Polyanetholsulfonate and Sodium Amylosulfate
Authors: Traub Walter H. , Fukushima Paula I.
Published: January 2020

Fresh, defibrinated human blood (80 vol%, i.e., 80% [vol/vol] of a 2-ml final assay volume) from two healthy adult donors killed “delayed serum-sensitive” (DSS) and “promptly serum-sensitive” (PSS) strains of
Serratia marcescens
, PSS control strain
Escherichia coli
C,
Bacillus subtilis
strain ATCC 6633, and
Micrococcus lysodeikticus
ATCC 4698 in a kinetic manner comparable to that of fresh human serum (80 vol%). However, heat-inactivated (56°C, 30 min), defibrinated human blood revealed markedly reduced or a total lack of β-lysin activity against the
B. subtilis
assay strain. Similarly, lysozyme activity of defibrinated blood was diminished somewhat by heat treatment, as determined with the
M. lysodeikticus
assay strain. Addition of 500 μg of sodium polyanetholsulfonate (SPS) per ml to 80 vol% of fresh, defibrinated human blood completely neutralized blood bactericidal activity against all assay strains of
S. marcescens, E. coli
C, and
B. subtilis
; however, SPS at this concentration failed to abolish lysozyme activity for prolonged periods of incubation. Addition of 500 μg of sodium amylosulfate (SAS) per ml to 80 vol% of fresh defibtinated human blood resulted in protection of cell inocula of DSS strains of
S. marcescens
only; SAS failed to protect cell inocula of the PSS strains of
S. marcescens, E. coli
C,
B. subtilis,
and
M. lysodeikticus
for extended periods of observation. Based on these data, it is recommended that blood culture specimens that are first drawn into specimen containers (such as Vacutainer tubes or the like) at the patient's bedside, and which contain ≥250 μg of SPS per ml, be diluted into suitable broth media with at least ≥250 μg of SPS per ml by the receiving laboratory within 2 to 4 h after procurement of the specimen. This procedure would ensure continued, adequate neutralization of the specimen's inherent β-lysin, lysozyme, and complement- and antibody-mediated bactericidal activities.
Radial immunodiffusion test with a Brucella polysaccharide antigen for differentiating infected from vaccinated cattle
Authors: Diaz R , Garatea P , Jones L M , Moriyon I
Published: January 2020

A Brucella antigen containing polysaccharide but lacking smooth lipopolysaccharide was employed in a rapid radial immunodiffusion test. With this serological test, cattle infected with Brucella abortus could be identified in recently vaccinated herds which had high numbers of reactors to standard diagnostic tests.
Evaluation of preservative fluid for urine collected for culture
Authors: Lauer B A , Reller L B , Mirrett S
Published: January 2020

Immediate culture or refrigeration of urine is recommended, but not always practical. Therefore, we evaluated the Becton-Dickinson Urine Culture Kit containing a boric acid-glycerol-sodium formate preservative in a study of 1,000 clinical urine specimens. Each specimen was cultured a total of four times by the surface streak technique with a 0.001-ml calibrated loop. After an initial reference culture (culture 1), a portion of urine was poured into a clean nonsterile paper cup, aspirated into a urine transport tube, and recultured immediately (culture 2). The original specimen cup was refrigerated for 18 to 24 h (culture 3), and the urine transport tube was held at room temperature for 18 to 24 h (culture 4) before repeat cultures. Eighty-eight of the initial reference cultures were positive (pure growth of greater than 10(5) bacteria per ml). Eighty-two (93.2%) of the 88 specimens positive on the reference culture were also positive after refrigeration or holding at room temperature in the transport tube for 24 h. There was one false-positive culture from refrigerated urine but none from the transport tube. Mixing of urine in a nonsterile container did not introduce detectable contamination. We conclude that the Becton-Dickinson Urine Culture Kit maintains a stable bacterial population in urine for up to 24 h as reliably as refrigeration. Urine for culture may be collected in a nonsterile container if it is immediately aspirated into the transport tube so that contaminants are not allowed to multiply.
Aeromonas primary wound infection of a diver in polluted waters
Authors: Joseph S W , Daily O P , Hunt W S , Seidler R J , Allen D A , Colwell R R
Published: January 2020

Two separate species of Aeromonas, A. sobria (not listed as a species in Bergey's Manual of Determinative Bacteriology, 8th ed.) and A. hydrophila, were primary pathogens isolated from the leg wound of a diver conducting operations in polluted waters. This is the first recorded instance of a primary infection of soft tissue in a human caused by two species of Aeromonas, one of which was resistant to tetracycline. Because of the very rapid development of this wound infection, cytotoxicity of these organisms was examined in several biological systems. A. sobria was hemolytic for sheep erythrocytes, cytotoxic for Y-1 adrenal cells, and enterotoxic in rabbit ligated intestinal loops, whereas A. hydrophila was hemolytic and cytotoxic. Pertinent clinical, bacteriological, and environmental features of the case are presented.
Distribution of hemolytic streptococci in respiratory specimens
Authors: Forrer C B , Ellner P D
Published: January 2020

One hundred thirty-seven isolates of beta-hemolytic streptococci were recovered from 623 pharyngeal cultures. Twenty-nine percent of these were group A, 10% were group B, 31% were group C, 11% were group F, 12% were group G, and 7% could not be grouped. The significance of non-group A isolates in pharyngitis could not be evaluated in the absence of viral and serological studies. Hemolytic streptococci were recovered from 9% of 799 lower respiratory cultures. All except one were non-group A, and other potential respiratory pathogens were also present in these specimens. It is our impression that the presence of hemolytic streptococci in lower respiratory tract specimens usually represents pharnygeal contamination.
Megasphaera elsdenii endocarditis
Authors: Brancaccio M , Legendre G G
Published: January 2020

A case of endocarditis caused by Megasphaera elsdenii is reported. This anaerobic grim-negative coccus has rarely been associated with human infections and has not previously been described as a cause of endocarditis.
Fluorescent antibody test kit for rapid detection and identification of members of the Bacteroides fragilis and Bacteroides melaninogenicus groups in clinical specimens
Authors: Holland R W , Stauffer L R , Altemeier W A
Published: January 2020

The Fluoretec fluorescent antibody test kit (Pfizer Inc., New York, N.Y.), developed for the rapid detection of members of the Bacteroides fragilis and B. melaninogenicus groups, was evaluated by testing 58 stock cultures and 76 clinical specimens. The test reagents detected 100% of 40 B. fragilis and B. thetaiotaomicron stock culture strains, although only 22% of 18 B. vulgatus, B. distasonis, and B. ovatus strains showed positive fluorescence. The 76 clinical specimens were evaluated by examining fluorescent antibody-stained smears of 49 specimens of purulent material and smears of 27 blood cultures which were positive for gram-negative bacilli by Gram stain or subculture. The fluoretec reagent detected members of the B. fragilis group in 28 (97%) of the 29 specimens of purulent material and all (100%) of the 16 blood cultures in which these anaerobes were demonstrated by culture. Overall, the Fluoretec reagent detected members of B. fragilis group in 44 (98%) of the 45 clinical specimens which were shown by culture to contain these anaerobes. Two of the 76 clinical specimens gave positive fluorescence for members of the B. fragilis group but failed to yield these organisms by culture. Members of the B. melaninogenicus group were detected by culture in 15 specimens and in each case their presence was demonstrated by the Fluoretec reagent. No members of the B. melaninogenicus group were isolated from five clinical specimens that gave positive fluorescence with the B. melaninogenicus reagent.
Reliable urease test for identification of mycobacteria
Authors: Steadham J E
Published: January 2020

A simple, modified formation for urea broth gave consistent, reliable results to aid in the differentiation of Mycobacterium species. In a study of 1,346 isolates representing 17 different species, tests read after 7 days were distinct and reproducible. The use of this test facilitates the identification of Mycobacterium species, such as M. scrofulaceum, the M. avium complex, the M. terrae complex, and M. triviale.
Merthiolate treatment of pathogenic fungi
Authors: Deighton F J , Hall N K , Larsh H W
Published: January 2020

The action of Merthiolate on the pathogenic yeasts Blastomyces, dermatitidis, Histoplasma capsulatum, and Sporothrix schenckii was compared to the effect of treatment with formaldehyde. Concentrations of 1:10,000 and 1:5,000 Merthiolate for three exposure times (24, 48, and 72 h) at 4 and 25 degrees C were tested on three media (brain heart infusion with and without blood, and modified Sabouraud agar). The effect of Merthiolate on these three yeasts was primarily fungistatic, with maximum effect using 1:5,000 Merthiolate at 25 degrees C for at least 48 h. Mycelial suspensions of B. dermatitidis, H. capsulatum, S. shenckii, and the yeast phase of Cryptococcus neoformans were susceptible to the 1:5,000 Merthiolate concentration after 24 h of treatment. The antifungal effect of Methiolate varies with species and growth phase of the fungus. Concentration, time of exposure, and temperature of incubation are important variables.
Obligate anaerobes in clinical veterinary practice
Authors: Hirsh D C , Biberstein E L , Jang S S
Published: January 2020

Clinical specimens obtained from domestic animals were examined to determine the relative prevalence of obligate anaerobic bacteria and the species represented. Of 3,167 samples cultured anaerobically as well as aerobically, 2,234 were bacteriologically positive. Of these positive samples, 583 (26%) contained species of obligate anaerobic bacteria in a total of 641 isolates. Most positive samples contained anaerobes admixed with aerobic species, although 6% of such samples yielded pure cultures of obligate anaerobes. The most common sites from which anaerobes were isolated were abscesses (32% of abscesses cultured contained species of obligate anaerobes), peritoneal exudates (24%), and pleural effusions (20%). Bacteroides melaninogenicus, Bacteroides spp., Peptostreptococcus anaerobius, and Bacteroides ruminicola accounted in the aggregate for approximately 50% of all anaerobic isolates. Bacteroides fragilis accounted for 1% of all the isolates, and members of the genus Clostridium accounted for 8%.
Comparison of various McCoy cell treatment procedures used for detection of Chlamydia trachomatis
Authors: Evans R T , Taylor-Robinson D
Published: January 2020

McCoy cells treated in six different ways, in addition to untreated cells, were compared to determine which gave rise to the largest number of Chlamydia trachomatis inclusions when tested with a laboratory-passaged strain. The same batch of cells was treated by irradiation, preinoculation exposure to cytochalasin B or 5-iodo-2'-deoxyuridine, and postinoculation exposure to cycloheximide, hydrocortisone, or emetine. Significantly more inclusions were always found in cells which had been treated with cycloheximide than in cells treated in any other way. Conversely, untreated McCoy cells always had significantly fewer inclusions than cells which had received some form of treatment. Similar results were obtained when cycloheximide-treated, irradiated, and untreated cells were inoculated with urethral specimens containing unpassaged organisms.
Identification of Candida lusitaniae as an opportunistic yeast in humans
Authors: Holzschu D L , Presley H L , Miranda M , Phaff H J
Published: January 2020

Four yeast strains, causally associated with infection in a patient with acute myelogenous leukemia, were identified by standard methods currently used in yeast taxonomy as representatives of Candida lusitania van Uden et do Carmo-Sousa. Because this species has not been recognized previously as an opportunistic yeast in humans, molecular taxonomic methods were applied to confirm its identity. The nuclear deoxyribonucleic acid (DNA) base composition of two clinical isolates was shown to be 45.1 mol% guanine plus cytosine as compared to 44.7 mol% guanine plus cytosine for the type strain of this species. DNA/DNA reassociation experiments revealed more than 95% complementarity between the DNAs from the clinical isolates and that of the type strain of C. lusitaniae, thus confirming their classification by conventional taxonomy. A key is provided to differentiate C. lusitaniae from two phenotypically similar Candida species.
Counterimmunoelectroosmophoresis for detection of neonatal calf diarrhea coronavirus: methodology and comparison with electron microscopy
Authors: Dea S , Roy R S , Begin M E
Published: January 2020

A counterimmunoelectroosmophoresis (CIE) technique is described for the detection of calf diarrhea coronavirus antigens in intestinal contents. The antibody reagent was prepared in rabbits against the Nebraska calf diarrhea coronavirus adapted to Vero cells and purified by density gradient centrifugation. The method was applied to intestinal contents of diarrheic and normal calves and compared with electron microscopy (EM). Calf coronavirus antigens were detected in intestinal contents of 44% (21/48) of the diarrheic calves and 24% (4/17) of the normal calves. Two precipitin lines could be observed in the majority of the positive samples. When compared with EM, CIE detected more positive animals. In only two cases (2/20) CIE was negative despite the visualization of coronavirus particles by EM.
Bactericidal antibody response to Pseudomonas aeruginosa by adults with urinary tract infections
Authors: Smalley D L , Ourth D D
Published: January 2020

In this investigation we found that adults with upper urinary tract infections caused by Pseudomonas aeruginosa produced serum antibodies with bactericidal activity against the bacterium. Seventeen of 20 infected adults showed bactericidal activity with a titer range of 1:10 to 1:10,000.
Formalinized Chlamydia trachomatis organisms as antigen in the micro-immunofluorescence test
Authors: Wang S P , Kuo C C , Grayston J T
Published: January 2020

Chlamydia trachomatis organisms grown in HeLa 229 cell cultures were purified and formalinized for use in the micro-immunofluorescence test. As test antigens, they were stable when stored unfrozen at 4 degrees C for a long period of time without loss of type specificity and sensitivity.
Demonstration of antibodies against Yersinia enterocolitica lipopolysaccharide in human sera by enzyme-linked immunosorbent assay
Authors: Gripenberg M , Nissinen A , Väisänen E , Linder E
Published: January 2020

Antibodies against Yersinia enterocolitica serotype O:3 lipopolysaccharide present in sera from patients with Yersinia infection were studied by using an enzyme-linked immunosorbent assay. Of the sera with significant bacterial agglutination titers against Y. enterocolitica type O:3, 86% contained anti-lipopolysaccharide antibodies of the immunoglobulin G class. With the sera of some patients, we demonstrated increasing anti-lipopolysaccharide antibody levels of immunoglobulin G class in spite of decreasing bacterial agglutination titers. The assay was specific for lipopolysaccharide from Y. enterocolitica type O:3, and in inhibition experiments lipopolysaccharide could be detected in amounts of greater than or equal to 0.5 micrograms/ml.
Microimmunodiffusion Test for Nocardiosis
Authors: Blumer Sharon O. , Kaufman Leo
Published: January 2020

Sera from 71 patients with culturally proven nocardiosis were tested for precipitins against a pool of
Nocardia asteroides
and
N. brasiliensis
culture filtrates and against antigens from the supernatant of homogenized
N. asteroides
cells. A human nocardiosis case serum was used as a reference. Sera from 56 of the 71 cases were reactive with either the culture filtrate antigen, the homogenate antigen, or both antigens, resulting in an overall sensitivity of 79%. Sera from 35 of the patients (49%) were positive with the homogenate antigens, and 28 (39%) showed bands of identity with the reference serum. Sera from 50 nocardiosis cases (70%) were positive with the pooled culture filtrate antigens, and 29 (41%) produced bands of identity with the reference serum. Of 89 sera from patients with various systemic mycotic diseases, tuberculosis, or actinomycosis, 24 (27%) were positive with the nocardial homogenate antigens and 4 (4.5%) showed precipitin bands of identity. Thirty-five of the 89 sera (42%) were positive with the nocardial culture filtrate antigens, and 6 (6.7%) showed bands of identity. The majority of sera demonstrating false-positive reactions were from tuberculosis and actinomycosis cases. One of seven sera from well individuals produced a precipitin band with the culture filtrate antigen, but this was not a band of identity with reference serum. These antigens did not distinguish antibodies from patients with
N. asteroides, N. brasiliensis,
or
N. cavia
infections.
Physiological characterization of nutritionally variant streptococci
Authors: Cooksey R C , Thompson F S , Facklam R R
Published: January 2020

Twenty-five isolates of nutritionally variant streptococci submitted to the Streptococcus Laboratory of the Center for Disease Control over a 2-year period were tested for growth requirements and for biochemical reactions. After they were recovered from storage in blood at -170 degrees C, all isolates grew within 48 h in both thioglycollate broth and Todd-Hewitt broth supplemented with 0.001% pyridoxal.HCl. They grew better in the latter, even though they all grew on unsupplemented infusion agar, anaerobe blood agar, and chopped meat-glucose medium. Biochemical patterns of the isolates resemble those of five viridans streptococcal species. Two isolates had patterns which did not resemble those of any viridans species. Biochemical reactions obtained with heart infusion broth base biochemicals and carbohydrate fermentation media compared favorably for an overall agreement rate of 86.5% for key tests. Lactic acid and acetic acid were the major fermentation products detected with gas-liquid chromatography.
Application of fluoroimmunoassay to cerebrospinal fluid immunoglobulin G and albumin
Authors: Stevens R W , Elmendorf D , Gourlay M , Stroebel E , Gaafar H A
Published: January 2020

Special solid-phase fluoroimmunoasssay protocols were used to measure the amount of immunoglobulin G (IgG) and albumin in 1,511 samples of cerebrospinal fluid. The fluoroimmunoassay is an inhibition test conducted on the surface of a plastic probe, whose fluorescence is inversely related to the concentration of spinal fluid IgG or albumin. A microcomputer interfaced with the fluorometer calculates the sample IgG or albumin from a calibration curve based on standard cerebrospinal fluid values. The test and interpretation take less than 90 min. Correlation coefficients for over 100 cerebrospinal fluid samples tested by both fluoroimmunoassay and radial immunodiffusion were: IgG, 0.90 (slope, 1.04); albumin, 0.95 (slope, 0.99). The within-run precision (coefficient of variation) was: IgG, 4.4%; albumin, 6.3%. Run-to-run precision on a midrange sample was: IgG, 7.7%, albumin, 11.7%. These findings establish the simplicity, speed, and precision of the modified fluoroimmunoassay system for specific cerebrospinal fluid proteins.
Direct hemagglutination technique for differentiating Bacteroides asaccharolyticus oral strains from nonoral strains
Authors: Slots J , Genco R J
Published: January 2020

A simple and economical method for differentiating Bacteroides asaccharolyticus of oral sources from nonoral sources is described. The present data indicate that oral strains of B. asaccharolyticus strongly agglutinate sheep erythrocytes, whereas isolates from various nonoral sites typically are devoid of hemagglutination activity. The direct hemagglutination test may aid in determining the source of B. asaccharolyticus present in an infection, and thus the procedure has potential value as a means of biotyping.
Neutralization test in influenza: use in individuals without hemagglutination inhibition antibody
Authors: Gross P A , Davis A E
Published: January 2020

In influenza immunization trials, patients who were seronegative by the hemagglutination inhibition test could be divided into two groups according to preimmunization levels of neutralizing (Neut) and neuraminidase inhibition (NI) antibodies. The group with low levels of Neut and NI antibodies usually developed adequate levels of hemagglutination inhibition antibody after one dose of vaccine, and the group with undetectable Neut and NI antibodies did not.
Comparison of Minitek and conventional methods for the biochemical characterization of oral streptococci
Authors: Setterstrom J A , Gross A , Stanko R S
Published: January 2020

Fifty-six strains of viridans streptococci were tested in the Minitek system (BBL Microbiology Systems, Cockeysville, Md.), and the results were compared with those obtained by conventional methods. An overall agreement of 98.9% was obtained when the Minitek tests were performed as follows. (i) All disks were incubated anaerobically for 48 h except for esculin and arginine, which required 5 to 7 days. (ii) The arginine disks were overlaid with 0.1 ml of sterile mineral oil even though incubated anaerobically. (iii) The Voges-Proskauer tests were performed under aerobic and anaerobic conditions. (iv) All tests for carbohydrate fermentation except for raffinose and salicin were read after the addition of 2 to 3 drops of 0.025% phenol red (pH 7.2). Of all the tests performed, only the fermentation of starch resulted in such poor agreement as to be considered unreliable. Results for inulin (unavailable in the Minitek system) were obtained by conventional methods.
Use of protein A-treated sera in unmasking herpes simplex virus type 1 (HSV-1) immunoglobulin A and identifying HSV-1 immunoglobulin G as the predominant neutralizing antibody
Authors: Ratner J J , Sanford B A , Smith K O
Published: January 2020

Treatment of human sera with protein A reduced the amounts of immunoglobulin G (IgG), IgA, and IgM detected by radial immunodiffusion. This treatment also decreased the amount of herpes-specific IgG and IgM detected by radioimmunoassay, whereas it increased and even unmasked the amount of herpes-specific IgA detected. Comparison of protein A-treated sera with untreated sera indicated that herpes simplex virus type 1 IgG was responsible for more than 92 to 99% of the serum neutralizing activity.
Charcoal-yeast extract agar: primary isolation medium for Legionella pneumophila
Authors: Feeley J C , Gibson R J , Gorman G W , Langford N C , Rasheed J K , Mackel D C , Baine W B
Published: January 2020

Charcoal-yeast extract agar is a new bacteriological medium that supports excellent growth of the Legionella pneumophila. It results from modifications made in an existing L. pneumophila medium, F-G agar. Yeast extract, instead of an acid hydrolysate of casein, serves as the protein source. Beef extractives and starch are not added. Activated charcoal (Norit A or Norit SG) is included at 0.20% (wt/vol). Comparison of charcoal-yeast extract and F-G agars showed that a greater number of colony-forming units of L. pneumophila was recovered from a standardized tissue inoculum on charcoal-yeast extract agar (4.35 x 10(6) colony-forming units) than on F-G agar (4.85 x 10(4) colony-forming units). Macroscopic colonies of L. pneumophila were visible on the new medium within 3 days, whereas 4 days of growth was required on F-G agar.
Differentiation of Peptococcus and Peptostreptococcus by gas-liquid chromatography of cellular fatty acids and metabolic products
Authors: Lambert M A , Armfield A Y
Published: January 2020

Gas-liquid chromatographic (GLC) profiles of cellular fatty acids and metabolic products were useful in identifying strains of Peptococcus saccharolyticus, Peptococcus asaccharolyticus, Peptostreptococcus anaerobius, Peptostreptococcus micros, and Streptococcus intermedius. The GLC results supported the recent taxonomic decision to transfer aerotolerant Peptostreptococcus species to the genus Streptococcus. Because inconsistencies in the results prevented our differentiating Peptococcus prevotii. Peptococcus magnus, and Peptococcus variabilis by GLC, additional strains will have to been examined. These GLC techniques are amenable to routine use; however, for interlaboratory results to be meaningful, the classification and nomenclature of the anaerobic gram-positive cocci should be standardized.
Effects of Blood on Blood Culture Medium
Authors: Beaman Kenneth D. , Kasten Bernard L. , Corlett Cynthia L. , Gavan Thomas L.
Published: January 2020

The morphological and biochemical changes that occur after inoculation of sterile blood into a blood culture medium (tryptic soy broth) with sodium polyanetholesulfonate and CO
2
were investigated. Cellular changes, pH, PCO
2
, and PO
2
were monitored and evaluated. Erythrocytes became crenated and developed precipitated hemoglobin inclusions within 4 h. The lymphocytes appeared morphologically intact at 24 h, and by 48 h a few cells had undergone transformation. Many neutrophils were vacuolated at 24 h. Neutrophils capable of phagocytizing
Staphylococcus aureus
were observed after 18 h of incubation. Identifiable eosinophiles were present on day 6 of the study. A decrease in PO
2
in the unvented bottles from 44.4 to 8 mm of Hg occurred by 24 h. PO
2
remained low for 6 days, after which a slight increase occurred. An increase in PO
2
in the vented bottle from 51 to 58 mm of Hg occurred by 24 h of incubation. In both the vented and unvented bottles the PCO
2
increased. This increase was markedly more rapid in the unvented bottle. From a pH of 7.06 a decrease occurred for the first 24 h after inoculation, with the pH stabilizing at 6.8 in the vented bottles and at 6.6 in the unvented bottles. The biochemical changes that occurred in the vented culture bottles stabilized more rapidly than those of the unvented bottles. Changes caused by the addition of sterile blood to a blood culture medium resulted in conditions which departed considerably from accepted optima for the isolation of clinically important microorganisms. The phagocytosis of organisms that occurred may also have reduced the yield.
Neutralizing antibody responses of humans and mice to vaccination with Venezuelan encephalitis (TC-83) virus
Authors: Fillis C A , Calisher C H
Published: January 2020

Laboratory workers were vaccinated with Venezuelan encephalitis virus, strain TC-83. After 2 to 3 years, they were bled, and their sera were tested for neutralizing antibody to all known Venezuelan encephalitis subtypes and varieties. The results indicated higher titers to epizootic than to enzootic Venezuelan encephalitis viruses and suggested that individuals vaccinated with TC-83 do not produce significant neutralizing antibody to heterologous subtypes. Mice vaccinated, bled, and tested in a similar manner produced much the same antibody profiles as did the humans and resisted challenge with all Venezuelan encephalitis viruses tested. In addition, the serum dilution plaque reduction neutralization test used was shown to be highly specific and adequate for diagnosis of infections with Venezuelan encephalitis viruses.