bluuebunny/crossref_metadata_2025 huggingface dataset:
Characteristics of yellow-pigmented nonfermentative bacilli (groups VE-1 and VE-2) encountered in clinical bacteriology
Authors: Gilardi G L , Hirschl S , Mandel M
Published: January 2020

The morphological and physiological characteristics of 20 strains of motile, gram-negative, yellow-pigmented oxidative bacilli (groups VE-1 and VE-2) isolated in clinical bacteriology are described. Electron micrographs demonstrate the polar multitrichous flagella of group VE-1 and polar monotrichous flagella of group VE-2. Data obtained from guanine plus cytosine ratio studies of 56.8% for VE-1 and 68.9% for VE-2 distinguish the two groups of bacteria.
Cellular fatty acids of Alcaligenes and Pseudomonas species isolated from clinical specimens
Authors: Dees S B , Moss C W
Published: January 2020

The cellular fatty acid composition of 25 clinical isolates of Alcaligenes and Pseudomonas was determined by gas-liquid chromatography (GLC). The GLC fatty acid profiles of three species of Pseudomonas were markedly different from those of Alcaligenes. The most significant differences were the presence and relative amounts of hydroxy, branched-chain, and cyclopropane fatty acids. One of the major fatty acids in A. faecalis was a 17-carbon cyclopropane (17 delta) acid, whereas a 15-carbon branched-chain acid (13-methyl tetradecanoate) characterized isolates of P. putrefaciens. The determination of these fatty acids by GLC provides a rapid and specific means of distinguishing clinical isolates of Pseudomonas and Alcaligenes.
Rapid conversion of Histoplasma capsulatum, Blastomyces dermatitidis and sporothrix schenckii in tissue culture
Authors: Hempel H , Goodman N L
Published: January 2020

A simple method for positive identification of Histoplasma capsulatum, Blastomyces dermatitidis, and Sporothrix schenckii is given. Primary tissue cultures of guinea pig peritoneal macrophage were inoculated with the mycelial phase of each organism and after 24 h the cells were stained and observed microscopically. The characteristic yeast phase could then be observed allowing for positive identification.
Radioimmunoassay for detection of antibodies to Epstein-Barr virus in human infectious mononucleosis serum specimens
Authors: Hutchinson H D , Ziegler D W , Feorino P M
Published: January 2020

A rapid microradioimmunoassay (RIA) technique was adapted for quantitatively measuring antibody titers to antigens occurring in Epstein-Barr virus (EBV)-infected lymphoid cells. In these experiments two EBV-infected cell lines, HR1K and EB-3, were used as antigen-positive cells and Molt-4 was used as the negative control cells. The antibody titers of sera from suspected infectious mononucleosis patients were compared by RIA and indirect fluorescent antibody (IFA) methods. As determined by each of the methods, 14 of 19 sera had positive antibody titers and the remainder of the sera had negative antibody titers. Thus, the two methods agreed completely in differentiating sera with antibodies to EBV antigens. To further evaluate the antibody specificity of the RIA, the antibody titers of paired sera, pre- or early infection and postinfection, from five confirmed infectious mononucleosis patients were determined by RIA and IFA. Seroconversion was demonstrated by both RIA and IFA for each of the patients. Thus, the sensitivity and specificity of the two procedures are about the same.
Comparative susceptibility of eleven mammalian cell lines to infection with trachoma organisms
Authors: Croy T R , Kuo C C , Wang S P
Published: January 2020

Eleven mammalian cell lines, HeLa 229, HeLa M, Hep-2, FT, BHK-21, Vero, MK-2, MPK, L-WO5A2, McCoy, and L-929, were tested for their susceptibility to infection with Trachoma strains TW-3 (type C, ocular origin) and UW-5 (type E, genital origin). All the cell layers were pretreated with diethylaminoethyl-dextran before inoculation of the organisms, and the inocula were centrifuged onto the cell layers. HeLa 229 was found to be the most sensitive to infection as determined by inclusion counts. The next most susceptible were cell lines MK-2, Hep-2, McCoy, and HeLa M, in that order. Infectivity in these cells ranged from 89 to 12% of that observed in HeLa 229. The remaining cell lines, BHK-21, L-WO5A2, L-929, Vero, MPK, and FT, were much less susceptible with infectivity less than 10% that of HeLa 229. HeLa 229 cells and 5-iodo-2'-deoxyuridine-pretreated McCoy cells have been used most extensively with Trachoma organisms in our laboratories. Infectivity in these two cell culture systems, both pretreated with diethylaminoethyl-dextran, was compared using 13 Trachoma strains of both ocular and genital origins of different immunotypes. The two systems performed similarly except with two type C, three type I, and one type J strains, With the type C, I and J strains tested, considerably fewer inclusions were found in 5-iodo-2'-deoxyuridine-pretreated McCoy than in HeLa 229 because inclusion formation of these strains in McCoy cells was not enhanced by 5-iodo-2'-deoxyuridine pretreatment.
Use of bile-esculin agar for rapid differentiation of Enterobacteriaceae
Authors: Lindell S S , Quinn P
Published: January 2020

Bile-esculin agar has been used for several years for the presumptive identification of group D streptococci. All members of the Enterobacteriaceae family will also grow on this medium, but only certain ones can hydrolyze esculin to 6,7-dihydroxycoumarin, which reacts with iron to produce a characteristic blackening of the medium. One thousand and six cultures from clinical specimens representing 20 genera were isolated and identified. Heavy inocula from fresh pure culture isolates on heart infusion agar were placed on bile-esculin agar slants and incubated at 35 C. The slants were examined at 4 h and again at 18 h for esculin hydrolysis. Shigella, Salmonella, Arizona, Proteus mirabilis, Proteus morganii, Providencia alcalifaciens, and Providencia stuartii all produced negative results. Klebsiella pneumoniae, Enterobacter aerogenes, Serratia marcescens, and Serratia rubidaea produced a positive reaction in 4 h. The other remaining eight genera exhibited varying results. The use of this medium in conjunction with triple sugar iron-lysine iron agar has been of great value in differentiating the Klebsiella-Enterobacter-Serratia group from other Enterobacteriaceae.
Inhibitory effect in vitro of sodium polyanethol sulfonate on the growth of Neisseria meningitidis
Authors: Eng J , Iveland H
Published: January 2020

The influence of 0.05% sodium polyanethol sulfonate on the growth of 24 strains of Neisseria meningitidis in broth medium was examined; several of the strains were markedly inhibited. A paper disk method was evolved for screening the sensitivity of various bacteria to sodium polyanethol sulfonate on solid medium; the highest sensitivity was observed in N. meningitidis and Neisseria gonorrhoeae. Zones of growth inhibition were also observed in a proportion of strains in certain species of gram-positive cocci, whereas all gram-negative bacilli were uniformly resistant. The implications of these observations for the routine use of sodium polyanethol sulfonate in blood culture media are discussed.
Physicochemical and serological characteristics of respiratory virus fluorescein-isothiocyanate conjugates for fluorescent-antibody diagnosis
Authors: Lyerla H C , Hierholzer J C
Published: January 2020

Fluorescein-isothiocyanate (FITC) conjugates were prepared by improved methods from standard reference antisera to influenza A and B, mumps, parainfluenza 1, 2, 3, and 4, herpesvirus, respiratory syncytial virus, and adenovirus hexon. The antisera, prepared in a variety of animals, were fractionated three times with selected optimal concentrations of ammonium sulfate and yielded gamma globulins of adequate purity for conjugation with FITC. Conjugates containing optimal fluorescein-to-protein ratios of between 5 and 10 were produced in 2 h by dialysis labeling. Serological titers of each antiserum and conjugate were determined by complement fixation, hemagglutination-inhibition, serum neutralization, and indirect hemagglutination tests where appropriate. When corrected for dilution, the serological titers of the FITC conjugates were identical to those of the starting antisera. The fluorescent-antibody staining titers correlated well with one of the serological parameters of the original serum. The conjugates stained homologous antigens specifically and were free of nonspecific staining at the working dilution. Undesired staining of host cells which was a problem with some of the conjugates produced from sera containing cellular antibodies was removed by absorption with packed cells. These physicochemical and serological findings were then used as a guide in preparing high quality reagents for fluorescent-antibody identification of respiratory viruses.
Complement fixation test as an aid in the diagnosis of farmer's lung
Authors: Marx J J , Motszko C , Wenzel F J
Published: January 2020

Complement-fixing antibodies to Micropolyspora faeni were measured in farmers and controls. Farmers with known or suspected farmer's lung showed significantly higher titers than control groups. Variations in titer were measured and correlated with the clinical disease.
Preparation of Rocky Mountain spotted fever vaccine suitable for human immunization
Authors: Kenyon R H , Pedersen C E
Published: January 2020

Rocky Mountain spotted fever vaccine was produced from rickettsiae grown in chicken embryo cells in roller bottle cultures. The rickettsiae were concentrated and purified by passage through a sucrose gradient and inactivated with formalin. This vaccine satisfactorily passed preinactivation and final container testing and is believed to be superior to the presently available yolk sac vaccine.
Construction of an interpretive pattern directory for the API 10 S kit and analysis of its diagnostic accuracy
Authors: Robertson E A , MacLowry J D
Published: January 2020

A directory of test patterns and their interpretations has been prepared for identification of Enterobacteriaceae by using the 11-test API 10 S kit. The diagnostic accuracy of the directory and kit were evaluated by using records of test results for 37,476 isolates studied with the 21-test API 20 Enteric kit. Analysis indicates that 96.9% of the isolates would have been correctly identified at the genus level and 95.9% at the species level by using only the subset of tests included in the API 10 S.
Serotyping of Pseudomonas aeruginosa isolates from patients with cystic fibrosis of the pancreas
Authors: Zierdt C H , Williams R L
Published: January 2020

Pseudomonas aeruginosa isolates (173) from 144 patients with cystic fibrosis (CF) of the pancreas in seven hospitals were serotyped with the agglutination systems of Homma (1974) and Fisher et al. (1969). The two systems were complementary. Strains from CF patients were much less likely to furnish a stable type on repetitive typing tests than strains from other patients. This was related to the frequent occurrence of mucoid P. aeruginosa strains. The 173 strains were divided among 11 Homma serotypes. A single Homma type (type 8) capable of mucoid growth comprised 104 (60%) CF strains. Eight serotypes were detected in 77 strains from 48 CF patients in one hospital; three strains were detected in one hospital CF unit; and two strains were detected in each of five hospital CF units. The CF serotype comprised from 50 to 93% of CF strians inthe seven hospitals. These P. aeruginosa strains dissociated in vivo as judged by mucoid and nonmucoid colonies on primary culture plates and continued to dissociate during subcultures. Both colony type were the same serotype. The tendency to regard colonial phenotypes (mucoid, nonmucoid, rough) as separate strians was erroneous. Repetitive typing with the two systems gave better results than a single system. The mucoid P. aeruginosa strain is probably spread from patient to patient, rather than acquiring its mucoid characteristic de novo in the CF patient. It is not known why the mucoid CF strain has a peculiar predilection for CF patients, nor why it generally loses the quality in culture but retains it indefinitely in the patient.
Anaerobic bag culture method
Authors: Rosenblatt J E , Stewart P R
Published: January 2020

In a new method of anaerobic culture, a transparent, gas-impermeable bag is used and the anaerobic environment is established with copper sulfate-saturated steel wool. An Alka-Seltzer tablet generates carbon dioxide. The agar plate surface can be inspected through the bag at any time without interrupting the anaerobic atmosphere or disturbing other specimens. Methylene blue indicator strips are completely reduced by 4 h after the bag is set up and have remained reduced for as long as 3 weeks. Growth of 16 different stock culture anaerobes was generally equivalent by the bag and GasPak jar methods. Yield and growth of anaerobic isolates also were equivalent with 7 of 10 clinical specimens; from the other 3 specimens, 13 isolates were recovered, 5 by both the bag and jar methods and the rest by one method or the other. No consistent differences were found between the anaerobic bag and GasPak jar methods in the yield of anaerobes from clinical specimens. Early growth (24 h of incubation) of anaerobes from one specimen was detected with the bag method.
Rapid presumptive bacteriological diagnosis of Legionnaires disease
Authors: Faine S , Edelstein P , Kirby B D , Finegold S M
Published: January 2020

A simple, relatively rapid silver impregnation stain has been found to stain Legionella pneumophila effectively in paraffin-embedded tissue sections while permitting visualization of histological detail. It may also be used to stain the organism in body fluids. The stain is not specific and thus must be confirmed by direct fluorescent-antibody technique or culture, but, in the absence of other bacilli demonstrable by Gram or other stain, visualization of typical bacillary forms in a patient with illness compatible with Legionnaires disease provides strong presumptive evidence supporting this diagnosis.
Effect of various histological fixatives on fluorescent antibody detection of Legionnaires disease bacteria
Authors: Thomason B M , Van Orden A , Chandler F W , Hicklin M D
Published: January 2020

Human lung tissue containing the Legionnaires disease bacterium was fixed in seven different histological fixatives, processed, and embedded in paraffin. Deparaffined sections from each were stained by fluorescent antibody and by Dieterle silver impregnation. With the fluorescent antibody stain, the Legionnaires disease bacterium could be detected in tissues prepared with any of the fixatures, but the Dieterle silver impregnation was not satisfactory on Zenker-fixed tissues.
Effect of enzymes on rotavirus infectivity
Authors: Barnett B B , Spendlove R S , Clark M L
Published: January 2020

The infectivity of a bovine rotavirus was enhanced 140-, 8-, and 3-fold, respectively, by trypsin, protease, and lactase. Ficin, carboxypeptidases A and B, lysozyme, and beta-galactosidase had little effect on the infectivity. Chymotrypsin caused a threefold decrease in the infectivity. Trypsin acts directly on the rotavirus and not on the host cell.
Neutralization of Human Serum β-Lysin by Sodium Polyanetholsulfonate and Sodium Amylosulfate
Authors: Traub Walter H. , Ima Paula I. Fukushi
Published: January 2020

Normal fresh and heat-inactivated (56°C, 30 min) human sera (80 vol%, i.e., 80% [vol/vol] of a 2-ml assay volume) killed
Bacillus subtilis
ATCC 6633 cell inocula of 1.5 × 10
4
colony-forming units per ml within 1 to 2 h after exposure. The
B. subtilis
assay strain proved slightly and reversibly susceptible to 5 μg of egg white lysozyme per ml. Seitz filtration of fresh human serum completely removed β-lysin activity; significant amounts of serum lysozyme were removed as well, as determined with the bioassay strain
Micrococcus lysodeikticus
ATCC 4698. However, bactericidal activity of human serum via classical or alternative complement pathway activation remained intact. Addition of 0.01 M dithiothreitol to fresh human serum abolished β-lysin activity, but not that of serum lysozyme. Chelation of fresh and heat-inactivated human serum with 0.01 M MgCl
2
+ 0.01 M ethylene glycol tetraacetic acid, but not with 0.01 M ethylenediaminetetraacetic acid, markedly retarded β-lysin activity; however, lysozyme activity remained unaffected. Chelation of serum with 0.01 M MgCl
2
+ 0.01 M ethylene glycol tetraacetic acid + 0.01 M CaCl
2
completely abrogated β-lysin activity, but not that of lysozyme. Absorption of human serum with 10 mg of bentonite per ml (10 min, 37°C) completely removed β-lysin and lysozyme activity, but failed to affect serum bactericidal activity against
Escherichia coli
control strain C. Reconstitution of 50 vol% of bentonite-absorbed serum with 40 vol% of heat-inactivated human serum restored both β-lysin and lysozyme activity. Addition of either 63 to 500 μg of sodium polyanetholsulfonate per ml or 63 to 500 μg of sodium amylosulfate per ml to 80 vol% of fresh human serum completely neutralized β-lysin activity for the entire observation period of 22 h.
Growth of Legionnaires disease bacterium (Legionella pneumophila) in chemically defined medium
Authors: Warren W J , Miller R D
Published: January 2020

A chemically defined medium containing 21 amino acids and inorganic salts was developed which supported the growth of four isolates of Legionnaires disease bacterium (Legionella pneumophila). Growth in liquid defined medium at 37 degrees C with shaking approximated the generation time and growth kinetics observed for growth in complex media. After a 3-h lag, the culture grew exponentially with a generation time of 6 h and reached a maximum optical density of 230 Klett units (170 Klett units corrected for pigment). A soluble brown pigment was first observed as the culture entered late exponential to early stationary phase of growth. Morphologically, L. pneumophila grew in the liquid defined medium with extensive filamentation and numerous intracellular lipid granuoles. L-Serine, L-methionine, and L-cysteine were required for optimum growth. The latter amino acid could be replaced by L-cystine or reduced glutathione but not by D-cysteine, thiomalate, thioglycollate, or 2-mercaptoethanol. Ferric iron was needed for maximum growth, but supplemental iron was not an essential growth requirement. Carbohydrates (i.e., glucose) or organic acids did not stimulate growth. In fact, pyruvate, acetate, and citrate all gave varying degrees of inhibition (69, 37, and 0% of control growth, respectively).
Serotyping of Serratia marcescens: current status of seven recently described flagellar (H) antigens
Authors: Traub W H , Fukushima P I
Published: January 2020

The slightly revised, current scheme of 20 flagellar (H) antigens of Serratia marcesens was examined. The seven new H antigens were demonstrated to be antigenically distinct as determined with Le Minor's H-immobilization test. The H-immobilization antibodies of rabbit anti-H immune sera proved resistant to treatment with 2-mercaptoethanol and dithiothreitol, respectively. On the other hand, dual absorptions of rabbit anti-H immune sera with killed cells of Staphylococcus aureus strain Cowan I, i.e., protein A, failed to reduce significantly H-immobilization titers of rabbit sera, although human immunoglobulins G and M were bound by protein A. It was tentatively concluded that the 2-mercaptoethanol- and dithiothreitol-refractory H-immobilizing rabbit antibodies belonged to the immunoglublin M class. H-antigen (phase) variation was not demonstrable in several extramural, clinical isolates of S. marcescens for which this phenomenon had been claimed. Rather, four of these six isolates were found to consist of cell populations of two distinct serotypes, as also borne out by bacteriocin typing; the flagellar H-antigens of the remaining two isolates were stable, with minor, hterologous H-antigen cross-reactivity.
Improved indirect hemagglutination test for cytomegalovirus using human O erythrocytes in lysine
Authors: Yeager A S
Published: January 2020

A modified indirect hemagglutination test for cytomegalovirus antibodies is described in which glutaraldehyde-fixed human O cells, rather than sheep cells, are used. Nonspecific hemagglutination was reduced by use of the optimal tannic acid concentration for each cell batch and the addition of 0.1 M lysine to the phosphate-buffered saline in which the fixed, tanned, sensitized cells were resuspended for use in the test. Three of 349 sera showed nonspecific hemagglutination by this technique. Antigen made by freezing phosphate-buffered saline (pH 7.2) over an infected monolayer can be used at dilutions of 1:8 to 1:15. Fixed, tanned, sensitized cells ready for use in the test can be stored in liquid nitrogen for up to 8 months. Use of cryoprotectants and washing after thawing is unnecessary. Simplification of the assay permits one person to screen 300 sera in 1 day or to determine the immune status of a potential donor or recipient in 45 min after the test is set up. The modified assay compares favorably in sensitivity with the complement fixation test and with previously described methods for performing the indirect hemagglutination test.
O antigen grouping of Morganella morganii (Proteus morganii) by slide agglutination
Authors: Penner J L , Hennessy J N
Published: January 2020

Antisera were prepared against Morganella morganii (Proteus morganii) type strains from the scheme described by Rauss et al. (Acta Microbiol. Acad. Sci. Hung. 22:315--321, 1975) and Raus and Vörös (Acta Microbiol. Acad. Sci. Hung. 6:233--248, 1959; Acta Microbiol. Acad. Sci. Hung. 14:195--198, 1967). The specificities of the somatic (O) antigens were studied using the passive hemagglutination and slide agglutination techniques. Previously unreported interstrain relations were observed, and O groups were provisionally defined on the basis of related strains. Of 143 isolates, collected mostly from two hospitals, 96% could be placed in one or another of the O groups. Forty-eight percent belonged to O group 1 and most of these were of two serotypes O1ab, 2 (20%) and O1ad, 2 (12%).
Serotyping of non-cholera vibrios
Authors: Smith H L
Published: January 2020

The serotyping system for non-cholera vibrios reported by us in 1965 has been re-evaluated and extended. Results from serotyping 2,624 cultures are presented. These isolates were accumulated over a period of 20 years and came from clinical and environmental samples in many parts of the world. Included is a discussion of the use of terms "Vibrio cholerae" and "non-cholera vibrios" for clinical reports.
Entamoeba histolytica: efficacy of microscopic, cultural, and serological techniques for laboratory diagnosis
Authors: Sheehan D J , Bottone E J , Pavletich K , Heath M C
Published: January 2020

Of 110 subjects with clinical evidence of amebiasis, 15 (14%) were shown to be infected with Entamoeba histolytica. Microscopic examination of stool specimens rendered a diagnosis in all eight cases of localized intestinal infection, but in only one of seven patients with invasive amebiasis. Culture was concomitantly diagnostic in six patients intestinal amebiasis and in one patient with extraintestinal infection. Assay for antibody to E. histolytica by counterimmunoelectrophoresis and indirect hemagglutination were each 100% effective in all cases of invasive amebiasis and in diagnosing two of eight patients with intestinal infection. Stool specimens of 15 patients revealing intestinal parasites other than E. histolytica failed to demonstrate cultural or serological evidence of amebiasis. Low levels of antibody were observed in the indirect hemagglutination assay in four patients with disease other than amebiasis and in three control sera positive for rheumatoid factor. By counterimmunoelectrophoresis, reactive sera were only encountered among those derived from patients with amebiasis. Six of seven patients with hepatic amebiasis may have gone undiagnosed if not for serology.
Use of a semiselective medium to culture Legionella pneumophila from contaminated lung specimens
Authors: Edelstein P H , Finegold S M
Published: January 2020

Legionella pneumophila was successfully isolated, using a semiselective medium, from two of three lung specimens heavily contaminated with other organisms. This medium is composed of charcoal yeast extract agar, supplemented with vancomycin and polymyxin B. L. pneumophila was observed at 8 days on plates containing less than or equal to 40 units of polymyxin B and less than or equal to 1 microgram of vancomycin per ml.
Clinical Evaluation of the MICRO-ID, API 20E, and Conventional Media Systems for Identification of
Enterobacteriaceae
Authors: Edberg Stephen C. , Atkinson Beverly , Chambers Carol , Moore M. Helen , Palumbo Lucy , Zorzon Corine F. , Singer Jacques M.
Published: January 2020

MICRO-ID (General Diagnostics, Morris Plains, N.J.) is a new kit system designed for the identification of
Enterobacteriaceae
in 4 h. It consists of 15 biochemical tests of paper disks. Each test is in its own compartment in a molded plastic tray. Only one reagent need be added to the system (2 drops of 20% KOH, which is added to the Voges-Proskauer test). Based on the pattern of positive and negative biochemical test results, a five-digit octal code number is calculated. An identification is derived from a computer-generated identification manual. A study was conducted to compare three systems—the MICRO-ID 4-h and the API 20E (Analytab Products Inc., Plainview, N.Y.) 18- to 24-h systems and a conventional media system—to measure the ability of each to identify members of the family
Enterobacteriaceae
. Comparison tables, rather than simple percentage agreement tables, were generated to define the particular strengths and weaknesses of each system and allow the laboratory to best use the data. The MICRO-ID compared quite favorably with conventional media. MICRO-ID yielded incorrect identifications with 1.5% of the isolates tested (API 20E, 4.7% misidentification rate). Half the MICRO-ID misidentifications occurred when the system identified a
Citrobacter diversus
as a lysine-negative
Escherichia coli
; all gave one octal number. A direct comparison of the MICRO-ID and API 20E was of limited value because percentage agreements were merely the sums of the errors of each. The ease of inoculation, the requirement for the addition of only one reagent, and the 4-h capability make the MICRO-ID system an extremely attractive development in the field of bacterial identification.
Medium for isolation of Yersinia enterocolitica
Authors: Dudley M V , Shotts E B
Published: January 2020

A new differential plate medium, CAL (cellobiose-arginine-lysine) agar, was developed and evaluated for the isolation of Yersinia enterocolitica. For all but two isolates tested, distinctive colonies were formed on CAL agar within 40 h of incubation at 25 degrees C. These colonies were readily recognized and isolated from a mixed inoculum containing 10 other bacteria commonly found in water and fecal specimens. Y. enterocolitica was presumptively identified based on colonial morphology from 9 of 10 fecal cultures containing the organism when inoculated to CAL agar and incubated for 40 h at 25 degrees C. No false-positive identification were made. The medium has been shown to be as sensitive as MacConkey agar in supporting growth of Y. enterocolitica, and it can be used in broth from for enumeration of the organism from water and liquid specimens. The medium is easily made and does not need to be autoclaved.
Antibody response to monovalent A/New Jersey/8/76 influenza vaccine in pregnant women
Authors: Murray D L , Imagawa D T , Okada D M , St Geme J W
Published: January 2020

The decision to implement a mass immunization program with A/New Jersey/8/76 (Hsw1N1) influenza vaccine provided a unique opportunity to evaluate immunological responses during pregnancy. Fifty-nine pregnant and 27 nonpregnant women participated in this study. Influenza virus hemagglutination-inhhibition antibody titers were determined to A/New Jersey/8/76 (Hsw1N1), A/Japan/305/57 (H2N2), and A/Hong Kong/8/68 (H3N2) before and after a single dose of monovalent (200 chick cell agglutination units) influenza A/New Jersey/8/76 (Hsw1N1) vaccine. The difference in titers between pregnant and nonpregnant women was insignificant. Treatment of the sera with 2-mercaptoethanol disclosed a similar immunoglobulin M response to the vaccine in both groups. The mean fold rise in heterologous antibody titer was similar in pregnant and nonpregnant women. This study demonstrated that pregnant women were able to respond to an original myxovirus antigen, influenza A/New Jersey/8/76, in a manner equivalent to nonpregnant, age-matched controls.
Solid-phase radioimmunoassay of serum immunoglobulin A antibodies to respiratory syncytial virus and adenovirus
Authors: Halonen P , Bennich H , Torfason E , Karlsson T , Ziola B , Matikainen M T , Hjertsson E , Wesslen T
Published: January 2020

A solid-phase radioimmunoassay for detecting respiratory syncytial virus and adenovirus serum immunoglobulin A (IgA) antibodies was developed. An antigen consisting of purified adenovirus type 2 hexons or a crude lysate of respiratory syncytial virus-infected cells was first adsorbed onto polystyrene beads. The coated beads were then incubated with dilutions of serum, and IgA antibodies which attached to the solid-phase virus antigen were subsequently detected with 125I-labeled anti-human alpha antibodies. The anti-human alpha antibodies used were isolated by immunosorbent chromatography from rabbit antiserum produced by immunization with IgA purified from serum of an IgA myeloma patient. A total of 46 serum specimens from 13 patients with respiratory syncytial virus infections and 10 patients with adenovirus infections were tested. Complement fixation, homologous IgG and IgM radioimmunoassay, and heterologous IgA radioimmunoassay testing were also done. Specific values higher than 10,000 cpm were often reached with convalescent serum specimens, and positive-to-negative serum binding ratios of 50 or more were frequently obtained with lower serum dilutions. IgA titers of convalescent sera were from 1,000 to 16,000, and with few exceptions a fourfold or greater rise in the IgA titer was detected in the homologous IgA radioimmunoassay.
Factors involved in enzyme-linked immunoassay of viruses and evaluation of the method for identification of enteroviruses
Authors: Herrmann J E , Hendry R M , Collins M F
Published: January 2020

A quantitative enzyme-linked immunosorbent assay was used for identification of selected enteroviruses: poliovirus type 1, echovirus type 6, coxsackievirus A type 9, and coxsackievirus B types 1 through 6. Partially purified viral antigens or virus-specific antibodies were adsorbed to polystyrene spectrophotometer cuvettes, which permitted the assays to be reported and compared in terms of enzyme units specifically reacting. Both the adsorbed antigen and the adsorbed antibody methods were approximately equal in terms of sensitivity and specificity of reaction. By use [14C]leucine-labeled enteroviruses, the amount of virus that bound to the plastics used was shown to be dependent on the purity of the virus preparation used, but it was higher than the amount that was bound by plastics coated with viral antibody. Diluents which contained 0.15% (vol/vol) Tween 20 and 2.0% (wt/vol) bovine serum albumin in phosphate-buffered saline, pH 7.2, were found to be the most effective in inhibiting nonspecific adsorption of immunoreagents. However, the presence of these inhibitors in phosphate-buffered saline solutions also caused desorption of virus or viral antibody during immunoassays; the amount of virus desorption varied with the type of preparation used, and antibody desorption was dependent on the concentration of antibody initially used for adsorption. For specific identification of a given enterovirus type by the enzyme-linked immunosorbent assay method used, approximately 10(5) plaque-forming units of virus per assay tube were required.
Differentiation of Pasteurella haemolytica biotypes A and T with growth inhibitors
Authors: Olmos A , Biberstein E L
Published: January 2020

Differentiation of type A and type T cultures of Pasteurella haemolytica was accomplished by paper disks containing 0.3 microgram of penicillin G, which produced zones of inhibition larger than 10 mm with type A but not with type T strains. Basic fuchsin (0.2 microgram/ml), brilliant green (0.005 microgram/ml), methylene blue (3.1 micrograms/ml) in brain heart infusion broth permitted the growth of type T but that of type A isolates.
Detection of adenovirus antibody by counterimmunoelectrophoresis
Authors: Petric M , Middleton P J , Mankikar S D
Published: January 2020

Counterimmunoelectrophoresis was adapted as a rapid, sensitive assay for detection of adenovirus group-specific antibody. This assay was six times more sensitive than the complement fixation test and was successfully adapted for use with microliter quantities of serum or whole blood.
Cell-mediated and humoral immune responses to herpes simplex virus and cytomegalovirus in renal transplant patients
Authors: Haahr S , Møller-Larsen A , Andersen H K , Spencer E S
Published: January 2020

Cell-mediated immunity to herpes simplex virus and cytomegalovirus, using the lymphocyte transformation test and interferon induction in lymphocytes, was studied in 59 patients from 1 day to 7 years after allotransplantation and compared with the results in normal subjects. Both parameters were permanently depressed with regard to cytomegalovirus. With herpes simplex virus, interferon production was also permanently depressed, whereas the transformation reaction was normal during the first year after transplantation and only slightly depressed in patients more than 1 year after transplantation. In 6 patients the above-mentioned assays and the complement fixation reaction were performed serially and related to the clinical signs of herpes simplex virus and cytomegalovirus infection. The relationship between depression of the transformation reaction and interferon production in lymphocytes and the occurrence of clinically evident herpes simplex virus and cytomegalovirus infections was, however, equivocal. The humoral immune response to herpes simplex virus was measured by the complement fixation test and the more sensitive antibody-dependent, cell-mediated cytotoxicity reaction, and a good correlation was found between these two tests, although only a few persons were found to be negative in the antibody-dependent, cell-mediated cytotoxicity reaction. The suggestion is made that only a few adults are "true" herpes simplex virus seronegative.
Rapid identification of Enterobacteriaceae with the micro-ID system versus API 20E and conventional media
Authors: Barry A L , Badal R E
Published: January 2020

The Micro-ID system for rapid (4 h) identification of Enterobacteriaceae was evaluated by testing 433 enteric bacilli and 9 other gram-negative bacilli. Each isolate was identified with conventional tubed media and was also tested in the Micro-ID and API 20E systems. The overall accuracy of both systems was 97%. Micro-ID tests for the Voges-Proskauer reaction, indole and H2S production, and ornithine and lysine decarboxylase all demonstrated a 97 to 99% correlation with conventional methods. Only 86% of the Micro-ID urease tests agreed with Christenson urea agar. Two inoculum densities were tested in Micro-ID panels, with 157 stock cultures. Over 90% of the tests were unaffected by changes in inoculum density. Tests with four control strains suggested that the Micro-ID system was more reproducible when a light inoculum was used. The Micro-ID system was found to be a very convenient method for rapid, accurate, and precise identification of the Enterobacteriaceae.
Practical technique for quantitating anaerobic bacteria in tissue specimens
Authors: Spengler M D , Rodeheaver G T , Edlich R F
Published: January 2020

Quantitative bacteriology has considerable influence on the care and management of surgical wounds. Heretofore, these techniques have been limited to measurements of aerobic bacteria. A technique is reported herein which permits quantitation of obligately anaerobic bacteria in tissue specimens. This technique is easily reproduced in any clinical laboratory and eliminates the need for expensive anaerobic chambers.
Enzyme-linked immunosorbent assay for detection of Streptococcus pneumoniae antigen
Authors: Harding S A , Scheld W M , McGowan M D , Sande M A
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of Streptococcus pneumoniae polysaccharide antigen in cerebrospinal fluid and serum. Sensitivity and specificity were determined for purified antigen preparations. Specificity was also evaluated in the rabbit meningitis model, and the sensitivity was compared to counterimmunoelectrophoresis, using the infected rabbits' cerebrospinal fluid and serum. The ELISA was a specific technique for detecting S. pneumoniae antigen. ELISA was 25 times more sensitive than counterimmunoelectrophoresis for purified antigen and resulted in an increased positivity of the cerebrospinal fluid and serum from infected rabbits. ELISA should prove very useful in the diagnosis of pneumococcal infections.
Assimilation of protocatechuic acid and p-hydroxybenzoic acid as an aid to laboratory identification of Candida parapsilosis and other medically important yeasts
Authors: Cooper B H , Land G A
Published: January 2020

Test for the ability of yeasts isolated from clinical specimens to utilize protocatechuic acid and p-hydroxybenzoic acid were carried out by using techniques that are commonly employed to test assimilation of carbon sources. A total of 60 isolates of Candida parapsilosis and 5 isolates of Candida humicola readily assimilated these two phenolic acids, whereas other Candida species gave uniformly negative results. Cryptococcus albidus, Cryptococcus terreus, and some isolates of Cryptococcus laurentii also assimilated protocatechuate and p-hydroxybenzoate, whereas Cryptococcus neoformans did not. Results of these tests suggest that assimilation of protocatechuate and p-hydroxybenzoate may be a useful characteristic, when used in conjunction with traditional tests, for identifying C. parapsilosis and C. albidus.
Reticulate bodies as single antigen in Chlamydia trachomatis serology with microimmunofluorescence
Authors: Yong E C , Chinn J S , Caldwell H D , Kuo C C
Published: January 2020

Formalin-fixed, purified reticulate bodies (RB) of Chlamydia trachomatis immunotype C/TW-3/OT were used as a serological test antigen in the microimmunofluorescence test. The sensitivity and specificity of the RB antigen were compared to elementary bodies (EB) used as antigens in the detection of C. trachomatis antibodies in human sera by microimmunofluorescence. RB reacted with all known C. trachomatis immunotypes with the same sensitivity as the homotypic EB. In routine serology with sera and endocervical secretions, the RB antigen had a sensitivity similar to that of the EB in detecting serum antibodies, endocervical secretion antibodies, and antibodies of immunoglobulin M and G classes. No false-positive reactions were detected with control sera. All positive reactions showed type-specific antibodies against an EB immunotype. RB seemed to demonstrate chlamydial group reactivity; sera from 10 psittacosis patients diagnosed clinically and serologically by complement fixation showed five positive, three equivocal, and two negative reactions. By immunofluorescence, RB appeared as distinct rings demonstrating uniform peripheral surface fluorescence at their rims. The EB appeared as pinpoint-sized dots. C/TW-3/OT RB used as a single test anitgen should provide a simple and sensitive serological assay for the detection of C. trachomatis antibody.
Pneumonia caused by Cunninghamella bertholletiae complicating chronic lymphatic leukemia
Authors: Kiehn T E , Edwards F , Armstrong D , Rosen P P , Weitzman I
Published: January 2020

A case of pneumonia caused by Cunninghamella bertholletiae is described in a patient with chronic lymphatic leukemia. The species of Cunninghamella, a genus in the order Mucorales, are characterized by the formation of conidia on the surface of an inflated conidiophore (vesicle). C. bertholletiae, not C. elegans, is the most appropriate name for this fungus. This zygomycete was resistant by in vitro sensitivity testing to amphotericin B. The source of the infection is not known.
CO2 requirement for growth of a Pasteurella strain isolated from rabbits
Authors: Targowski S , Targowski H
Published: January 2020

A Pasteurella strain with unusual growth requirements was isolated from a rabbit abscess. Copious growth of this organism was observed only on media supplemented with serum and incubated in the candle jar. Under anaerobic conditions, growth was moderate on enriched media. The organism did not grow on unsupplemented media under these conditions or in an aerobic environment on enriched media.
Cellular fatty acid composition of WIGA, a rickettsia-like agent similar to the Legionnaires disease bacterium
Authors: Moss C W , Dees S B
Published: January 2020

The cellular fatty acid composition of OLDA, a rickettsia-like agent isolated in 1947, was essentially identical to that of the Legionnaires disease bacterium (LDB). WIGA, another rickettsia-like agent isolated in 1959, contained the same fatty acids as OLDA and other LDB but differed significantly from these strains in relative amounts of the major acids present. The major acid of OLDA and other LDB was i-16:0 whereas a-15:0 was the major acid of WIGA.
Adaptation of a Microdilution Procedure to the
Limulus
Lysate Assay for Endotoxin
Authors: Prior Richard B. , Spagna Vincent A.
Published: January 2020

Endpoint determinations of endotoxin by the
Limulus
amoebocyte lysate assay have been facilitated by use of a microdilution procedure.
Rapid Identification of
Enterobacteriaceae
by Using Noncommercial Micro-Tests in Conjunction with API 20E Profile Data
Authors: George Sarah , Davis G. H. G.
Published: January 2020

A total of 371 (97%) of 382 strains of
Enterobacteriaceae
representing 26 species were identified cheaply, accurately, and rapidly by using a test system based upon standard 96-well microdilution plates and the API 20E profile data.
Periurethral anaerobic microflora of healthy girls
Authors: Bollgren I , Källenius G , Nord C E , Winberg J
Published: January 2020

The periurethral anaerobic and aerobic microfloras were investigated in 18 healthy premenarcheal girls, 5 to 14 years of age, by using a quantitative sampling method. Colonization of the female periurethral area with enterobacteria seems to be an important step in the development of urinary tract infections, and the present study was undertaken as a stage in elucidating factors that might control the establishment of urinary tract pathogens periurethrally. The study showed that obligate anaerobic bacteria constituted 95.0% (standard error, +/- 5.8%) of the total colony-forming units per square centimeter of periurethral area. An average of 7.0 different anaerobic and 2.7 different aerobic strains per specimen was obtained. The flora was dominated by anaerobic gram-positive cocci and gram-positive rods, whereas anaerobic gram-negative rods comprised a minor part. The most commonly encountered anaerobic isolates were peptococci and peptostreptococci, propionibacteria, bifidobacteria, eubacteria, and bacteroides in decreasing order of frequency. The aerobic flora consisted most commonly of nonhemolytic streptococci and diphtheroids. The findings suggest that the periurethral microenvironment is a distinctive ecological niche, separate from the fecal and skin biotas, although it has some characteristics in common with the vaginal flora.
Serodiagnosis of ram epididymitis by counterimmunoelectrophoresis, using Brucella ovis surface R antigen
Authors: Myers D M , Varela-Diaz V M
Published: January 2020

The counterimmunoelectrophoresis (CIE) technique was developed as a diagnostic procedure for ram epididymitis caused by Brucella ovis. CIE test results with sera from naturally and experimentally infected male sheep compared favorably with those obtained by gel diffusion and complement fixation employing the same B. ovis surface R antigen. The main advantage of CIE over gel diffusion consists in a significant reduction of the time required to detect precipitin formation, whereas both methods obviate several of the difficulties encountered with complement fixation tests for B. ovis.
Hemagglutination by
Escherichia coli
in Septicemia and Urinary Tract Infections
Authors: Ljungh Å. , Faris A. , Wadström T.
Published: January 2020

The agglutination of erythrocytes from various animal species by
Escherichia coli
was studied. The 405 strains of
E. coli
were isolated from urine in patients with urinary tract infections, from blood in septicemic patients, or from feces in persons without intestinal or urinary disorders. In urinary tract infections,
d
-mannose-resistant agglutination (MRHA) of human erythrocytes was the most common finding (23% of the strains). The highest frequency of mannose-sensitive hemagglutination (MSHA) attributed to type I (common type) pili occurred with guinea pig erythrocytes (11.5%). Of the 78
E. coli
strains isolated from blood cultures, 11 (14%) produced MRHA of human erythrocytes and only one gave MSHA. In the stool cultures, only 1 of 170
E. coli
strains was MSHA reacting, whereas 28 strains (16.5%) showed MRHA of human erythrocytes. No MRHA strain reacted with antiserum against colonization factor antigen (CFA)/I of pilus nature in enterotoxigenic human
E. coli
strains (O78:H12). MRHA of bovine erythrocytes, reputedly typical of enterotoxigenic
E. coli
of serogroups O6 and O8, was shown by only two strains, neither of which agglutinated with CFA/II antiserum. The most common hemagglutinating pattern of
E. coli
from urine and blood thus was MRHA for human erythrocytes. This agglutination may have been caused by pili or other surface properties of one or more serotypes. These may represent a new class of colonization-promoting antigens (adhesins).
Rapid differentiation of Streptomyces from Nocardia by liquid chromatography
Authors: Tisdall P A , Anhalt J P
Published: January 2020

A liquid chromatographic method is described for analysis of aerobic actinomycetes for isomers of diaminopimelic acid. One or two colonies of organism were hydrolyzed with 6.0 mol of HCl per liter at 121 degrees C for 15 min. The hydrolysate was neutralized and buffered with an NaOH solution (3 mol/liter) containing 0.15 mol of sodium borate per liter. Precolumn derivatization with dansyl chloride was used to form a fluorescent product for detection. Analysis was performed by reversed-phase, ion-pair chromatography. The L-diaminopimelic acid isomer was detected in all 10 strains of Streptomyces tested, and the meso-diaminopimelic acid isomer was detected in all 10 strains of Nocardia tested. Liquid chromatography was compared simultaneously with thin-layer chromatography in the analysis of three strains of aerobic actinomycetes. Liquid chromatography required less growth of the organisms, and analysis was completed within 1 h, compared with the 3 to 5 days required by thin-layer chromatography.
Evaluation of Micro-Media Quad Panels for identification of the Enterobacteriaceae
Authors: Kelly S A , Washington J A
Published: January 2020

The Micro-Media Quad Enteric Panel and coding system were evaluated by comparing the identifications of 193 stock and 216 clinical isolates, representing 31 species of Enterobacteriaceae, with those provided by conventional methods. The results corresponded for 91% of the organisms if only the single or most probable identification was accepted and for 93% if the correct identification was listed as one of several possible, but not the most probable, identification by the coding system. With 73% of the isolates, the single identification provided by the coding system was correct, and in 20% of instances the correct identification was listed as the first of several choices in descending order of probability but requiring additional tests for differentiation. In most of the latter instances, these additional tests consisted of serological confirmation of salmonellae or shigellae. Discrepancies consisted of incorrect identifications (4%) and organisms with code numbers not appearing in the coding system (3%).