bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of atypical aerobic gram-negative cocci isolated from humans
Authors: Hoke C , Vedros N A
Published: January 2020

Six atypical aerobic gram-negative cocci, isolated from humans, were classified taxonomically by genetic relatedness (DNA base pairs and DNA homology), fatty acid content, pigment absorption profile, and other physiological and biochemical characteristics. One strain (M448), which produced acid only from glucose, was identified as Neisseria meningitidis; one strain (CH), which produced acid from glucose, maltose, and sucrose, was identified as N. gonorrhoea; Branhamella catarrhalis was confirmed in a case of acute meningitis; N. canis was identified in a cat bite wound; and representative isolates of a proposed new pigmented species were found to have genetic affinities with N. meningitidis and N. perflava.
Identification of viridans streptococci isolated from clinical specimens
Authors: Ruoff K L , Kunz L J
Published: January 2020

Of 532 strains of viridans streptococci isolated from clinical specimens, 517 were identified by using a scheme based on the work of Facklam (R. R. Facklam, J. Clin. Microbiol. 5:184-201, 1977). The strains were distributed among nine of the species described by Facklam and an additional group of physiologically homogeneous streptococci not recognized in Facklam's scheme. The method for identification involves a battery of 10 tests that employ both conventional media and commercially prepared disks. The identification of most of the isolates was accomplished within 48 h.
Evaluation of a solid-phase immunoassay with fluorescein isothiocyanate-conjugated heterogeneous or monoclonal antibodies for identification of virus isolates, with influenza virus as a model
Authors: Phillips D J , Galland G G , Reimer C B , Kendal A P
Published: January 2020

A solid-phase immunofluorescence assay was evaluated for the identification of viruses isolated in tissue culture, with influenza virus as a model. Purified immunoglobulin G (IgG) from hyperimmune rabbit sera specific for contemporary strains of influenza A or B was covalently attached to microscopic plastic beads to capture virus. Fluorescein isothiocyanate (FITC)-conjugated antibodies of different specificities were then reacted with bound antigen, and the resulting complexes were quantified in a suitable filter fluorometer. The assay, with appropriately absorbed FITC-conjugated second antibody, reliably identified virus present in harvests from cell cultures infected with clinical specimens. For influenza A (H1N1) virus, sensitivity of detecting antigen was about 8- to 32-fold less when an FITC-conjugated monoclonal Igg antibody pool specific for epitopes in three different antigenic sites on influenza hemagglutinin was used as the second antibody as compared to when IgG from hyperimmune sera specific for virus or its components was used as the second antibody. The immunofluorometric assay provides a method for quantitative detection of viral antigen in tissue culture fluids and objective identification of virus type and subtype with FITC-conjugated reagents.
Infectivity of Chlamydia trachomatis in tissue culture with newborn calf serum
Authors: LaScolea L J , Baldigo S M
Published: January 2020

By using representative strains of Chlamydia trachomatis serotypes, the efficacy of substituting fetal bovine serum with newborn calf serum for the isolation of this organism in tissue culture was examined. The criteria used were the quality and quantity of the iodine-stained cytoplasmic inclusions and the characteristics of the McCoy cells. Complete substitution of fetal bovine serum with newborn calf serum produced a detrimental change in the quality of the cytoplasmic inclusions and a dramatic decline in the inclusion count (P less than 0.001) with all of the chlamydial strains tested. There appeared to be no significant alteration in the characteristics of the McCoy cells. It is recommended from this preliminary investigation that newborn calf serum should not be used for the isolation of C. trachomatis in tissue culture.
Postoperative infection caused by an unusual serotype of Streptococcus pneumoniae associated with multiple drug resistance
Authors: O'Donnell E D , Alter K E , Frenkel L D
Published: January 2020

A 15-month-old child developed an infectious pulmonary complication of open heart surgery. Cultures of the respiratory secretions showed growth of a 9L serotype of Streptococcus pneumoniae which was resistant to penicillin, tetracycline, and chloramphenicol. There was no evidence that the organism was spread among the family of the patient or hospital personnel.
Radioimmunoassay for detection of antibodies to Neisseria gonorrhoeae
Authors: Usategui M , Savard E V , Mondabaugh S M , Keigher N L
Published: January 2020

A radioimmunoassay has been developed and evaluated for the serological diagnosis of gonorrhea. Purified gonococcal antigen was obtained from a culture of Neisseria gonorrhoeae (B370) and labeled with 125I for use in a double-antibody test system. The test was evaluated in populations segregated by sex and risk. The specificity of the assay in females was 90.2% (55/61) in low risk, 82.2% (2,245/ 2,732) in medium risk, and 54.1% (335/619) in high risk. The sensitivity was 69% (20/29) in medium risk and 78.3% (288/367) in high risk. In males, test specificity was 92.3% (24/26) in low risk and 50% (48/96) in high risk. The sensitivity was 70.8% (143/202) in the high-risk group. The data in this study indicate that this assay should not be employed for screening of either high- or medium-risk populations.
Glucose-nonfermenting Gram-negative bacilli associated with clinical veterinary specimens
Authors: Mathewson J J , Simpson R B
Published: January 2020

Glucose-nonfermenting gram-negative bacilli (NFB) have been recognized recently as opportunistic pathogens of humans. With few exceptions, strains of NFB have not been considered important enough to be identified when isolated from animals. In this study, all NFB isolated during a 1-year period in a clinical veterinary microbiology laboratory were identified to determine their prevalence. Of the 347 strains of NFB obtained, the most common species were Pseudomonas aeruginosa, Acinetobacter calcoaceticus, Bordetella bronchiseptica, and Pseudomonas pseudoalcaligenes. Of all clinical veterinary specimens submitted for cultures, 10% contained nonfermenters.
Fractionation of Pneumocystis carinii antigens used in an enzyme-linked immunosorbent assay for antibodies and in the production of antiserum for detecting Pneumocystis carinii antigenemia
Authors: Maddison S E , Hayes G V , Ivey M H , Tsang V C , Slemenda S B , Norman L G
Published: January 2020

Cyst-rich suspensions of Pneumocystis carinii were obtained by differential and gradient centrifugation from heavily infected rat lungs. After preparation of an aqueous-soluble extract of the cyst-rich material, the insoluble residue was extracted with 8 M urea. Small amounts of infected human lung tissue and uninfected rat and human lung were processed similarly. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that both human and rat infected lung extracts contained a large protein (greater than 200,000 daltons). This component was not present in extracts of uninfected lung. In addition, an HCl-soluble extract was prepared from the cyst-rich suspension from infected rat lung. The urea-extracted antigen was most reactive in an enzyme-linked immunosorbent assay. Rabbit antiserum against the HCl-soluble antigen detected circulating antigen in patients' sera in a counterimmunoelectrophoresis assay.
Isoprenoid quinones of the genus Legionella
Authors: Karr D E , Bibb W F , Moss C W
Published: January 2020

Representative strains of each of the named species of Legionella were examined for isoprenoid quinones by reverse-phase thin-layer chromatography. All strains contained three or more ubiquinones (Q9, Q10, Q11, Q12, Q13) which were useful for placing the species into one of three distinct groups. Group 1 contained L. longbeachae, L. bozemanii, L. dumoffi, and L. gormanii; group 2 contained only L. micdadei; and group 3 contained only L. pneumophila. The identities of the quinones were established by UV spectroscopy and mass spectrometry.
Gas-liquid chromatography of positive blood cultures for rapid presumptive diagnosis of anaerobic bacteremia
Authors: Edson R S , Rosenblatt J E , Washington J A , Stewart J B
Published: January 2020

Gas-liquid chromatographic analysis of volatile fatty acid patterns was performed when growth was initially detected in 87 positive blood cultures. Isovaleric acid, butyric acid, or both were found in 39 of 43 (90%) blood cultures that contained anaerobes but were absent in all 44 cultures that contained facultatively anaerobic or aerobic bacteria. The detection of isovaleric acid indicated the presence of Bacteroides fragilis, as this acid was found in 88% of unimicrobial and 75% of polymicrobial bacteremias due to B. fragilis. Butyric acid was detected most often in blood cultures which grew Fusobacterium spp. (three of three), Clostridium spp. (six of seven), or both (three of four).
30 years of campylobacters: biochemical characteristics and a biotyping proposal for Campylobacter jejuni
Authors: Hébert G A , Hollis D G , Weaver R E , Lambert M A , Blaser M J , Moss C W
Published: January 2020

Several biochemical test systems were studied for their potential usefulness for the examination of strains of Campylobacter species. Most (81%) of the C. jejuni strains hydrolyzed sodium hippurate, but strains of C. fetus, C. sputorum, and C. fecalis did not. Some (46%) of the C. jejuni strains and all of the C. sputorum subsp. sputorum, C. sputorum subsp. bubulus, and C. fecalis strains hydrolyzed DNA, but the C. fetus and C. sputorum subsp. mucosalis strains did not. Strains of all species of Campylobacter grew on charcoal-yeast extract agar, but 47% of the C. jejuni strains did not. Alkaline phosphatase activity was recorded for some strains of C. jejuni, but all other species were negative for this activity. Aryl sulfatase activity was detected in 7% of the C. jejuni, 15% of the C. fetus subsp. fetus, and all of the C. sputorum subsp. sputorum, C. sputorum subsp. bubulus, and C. fecalis strains, but it was not detected in the C. fetus subsp. venerealis and C. sputorum subsp. mucosalis strains. Most (93%) of the C. jejuni but none of the other Campylobacter strains contained lactobacillic acid when examined for cellular fatty acids. On the basis of results from three of these tests (hippurate hydrolysis, DNA hydrolysis, and growth on charcoal-yeast extract agar), clinical strains of C. jejuni were placed in eight biotypes.
Hemagglutination by
Bordetella bronchiseptica
Authors: Bemis David A. , Plotkin Balbina J.
Published: January 2020

A total of 53 isolates of
Bordetella bronchiseptica
from dogs and pigs were tested for their ability to agglutinate chicken, horse, sheep, dog, pig, and guinea pig erythrocytes. No differences in hemagglutinating activity were attributed to the animal origin of the bordetella isolates. Horse and dog erythrocytes consistently resulted in the strongest hemagglutination reactions, whereas only 4% of the
B. bronchiseptica
isolates produced weak agglutination of chicken erythrocytes. A total of 85% of the isolates agglutinated horse, sheep, dog, pig, and guinea pig erythrocytes. One canine isolate with hemagglutinating activity, strain 110H, was examined to determine the nature of the hemagglutinin(s) involved. Hemagglutination was always accompanied by hemadsorption, as determined by dark-field or phase-contrast microscopy. Treatment of cells and cell extracts with heat or protease K inhibited the hemagglutination reaction. Sonicated bacterial cells had a greater hemagglutinating ability than did unsonicated live bacteria. The hemagglutination reaction was not inhibited by any of 17 sugars nor by
N
- acetylglucosamine or ethylene glycol-bis-(β-aminoethyl ether)-
N
,
N
-tetraacetic acid. Hemagglutinins were not detected in sonic extracts nor in several bacterial subunit fractions, including isolated pili. Antigens in some of these preparations were, however, detectable by indirect hemagglutination with anti-
B. bronchiseptica
serum. Isolated pili could not be detected on the erythrocyte surface by electron microscopy; however, serial sections of erythrocytes agglutinated by the live
Bordetella
organisms showed that the bacterial outer membrane and the erythrocyte surface were separated by a space of approximately 20 nm. This study provided additional circumstantial evidence that
B. bronchiseptica
pili or at least heat-labile surface proteins which extend some distance from the bacterial surface are involved in hemagglutination. Multiple hemagglutinins are likely to exist within this species since one isolate lacking pili also agglutinated canine erthyrocytes. The hemagglutinins of
B. bronchiseptica
need to be isolated and characterized before the hemagglutination reaction can be applied to studies of attachment.
Escherichia vulneris
: a New Species of
Enterobacteriaceae
Associated with Human Wounds
Authors: Brenner Don J. , McWhorter Alma C. , Knutson Jean K. Leete , Steigerwalt Arnold G.
Published: January 2020

The name
Escherichia vulneris
sp. nov. (formerly called Alma group 1 and Enteric group 1 by the Centers for Disease Control and API group 2 by Analytab Products, Inc.) is proposed for a group of isolates from the United States and Canada, 74% of which were from human wounds.
E. vulneris
is a gram-negative, oxidase-negative, fermentative, motile rod with the characteristics of the family
Enterobacteriaceae.
Biochemical reactions characteristic of 61
E. vulneris
strains were positive tests for methyl red, malonate, and lysine decarboxylase; a delayed positive test for arginine dihydrolase; acid production from
d
-mannitol,
l
-arabinose, raffinose,
l
-rhamnose,
d
-xylose, trehalose, cellobiose, and melibiose; negative tests for Voges-Proskauer, indole, urea, H
2
S, citrate, ornithine decarboxylase, phenylalanine deaminase, and DNase; and no acid from dulcitol, adonitol,
myo
-inositol, and
d
-sorbitol. Two-thirds of the strains produced yellow pigment. Most strains gave negative or delayed positive reactions in tests for lactose, sucrose, and KCN. The
E. vulneris
strains tested were resistant to penicillin and clindamycin, were resistant or showed intermediate zones of inhibition to carbenicillin and erythromycin, and were susceptible to 14 other antibiotics. DNA relatedness of 15
E. vulneris
strains to the type strain averaged 75% in reactions at 60°C and 69% in reactions at 75°C, indicating that they comprise a separate species. DNA relatedness to other species in the family
Enterobacteriaceae
was 6 to 39%, an indication that this new species belongs in the family.
E. vulneris
showed the highest relatedness to species of
Escherichia
(25 to 39%) and
Enterobacter
(24 to 35%). On the basis of biochemical similarity, the new species was placed in the genus
Escherichia.
The type strain of
E. vulneris
is ATCC 33821 (CDC 875-72).
Osteomyelitis Caused by
Moraxella osloensis
Authors: Sugarman Barrett , Clarridge Jill
Published: January 2020

Moraxella osloensis
osteomyelitis of the femur developed in a paraplegic man. He responded to treatment with oral ampicillin. Disease in humans caused by this unusual clinical isolate is reviewed.
Differential Effects of
Clostridium difficile
Toxins on Tissue-Cultured Cells
Authors: Donta Sam T. , Sullivan Nadine , Wilkins Tracy D.
Published: January 2020

Two immunologically distinct
Clostridium difficile
toxins elicited similar morphological changes on cultured cells, although there were differences in both toxin potency and cell sensitivities.
Use of potato flakes agar in clinical mycology
Authors: Rinaldi M G
Published: January 2020

A formulation comprised of commercially available potato flakes, glucose, and agar provides a medium which is inexpensive, is easily prepared, and is particularly useful for the identification of mold fungi encountered in the clinical mycology laboratory. The inclusion of appropriate antimicrobial agents results in a selective medium effective in the isolation and identification of medically important fungi.
Evaluation of three differential media for detection of Enterobacter agglomerans (Erwinia herbicola)
Authors: Bucher C , von Graevenitz A
Published: January 2020

Dextrin-fuchsine-sulfite medium (DFS), Rimler-Shotts agar (RS), and a new lysine-ornithine-mannitol agar (LOM) were tested for detection of Enterobacter agglomerans. In human stools, LOM and DFS were most sensitive at coliform-to-E. agglomerans ratios of less than or equal to 10(2) and E. agglomerans inocula of greater than 10(2) per plate. Both LOM and DFS detected one strain in 254 stools, but RS proved to be inhibitory.
Rapid detection in spinal fluid of beta-lactamase produced by ampicillin-resistant Haemophilus influenzae
Authors: Boughton W H
Published: January 2020

The chromogenic cephalosporin nitrocefin was used to detect the presence of beta-lactamase in cerebrospinal fluid of patients suffering from ampicillin-resistant Haemophilus influenzae meningitis. Five samples of spinal fluid containing ampicillin-resistant isolates were studied, and all had beta-lactamase activity. When samples of spinal fluid containing 33 ampicillin-sensitive isolates and 234 sterile specimens were tested, no beta-lactamase activity was detected.
Comparison of paired whole milk and dried filter paper samples for anti-enterotoxin and anti-rotavirus activities
Authors: Brown S E , Sauer K T , Nations-Shields M , Shields D S , Araujo J G , Guerrant R L
Published: January 2020

Milk specimens, 75 from cows immunized against cholera toxin and 35 from a human population in which enterotoxigenic Escherichia coli and rotaviral infections are endemic, were collected as paired filter paper and frozen whole milk samples. Each pair was tested for antibody activity against heat-labile E. coli and Vibrio cholerae enterotoxins. Additionally, 12 of the 35 paired human milk samples stored as frozen whole milk and dried on filter paper were tested for anti-rotavirus immunoglobulin A. Anti-enterotoxin and anti-rotavirus immunoglobulin A titers in milk dried on filter paper compared favorably with those of their frozen whole milk pairs. Filter paper samples offered considerable advantages for field collection, transportation, and storage over frozen liquid samples.
Elevated mannose levels detected by gas-liquid chromatography in hydrolysates of serum from rats and humans with candidiasis
Authors: Marier R L , Milligan E , Fan Y D
Published: January 2020

We have developed a gas-liquid chromatographic method for identification of mannose in serum hydrolysates by utilizing peracetylated aldononitrile derivatives. Experimentally infected rats and human subjects with candidiasis were studied. Peaks in unknown samples were identified by co-chromatography of reference carbohydrates. Inositol was used as an internal standard. Mannose was identified in hydrolysates of normal rat (391.85 +/- 66.18 micrograms/ml) and human (297.87 +/- 77.81 micrograms/ml) sera. Significantly increased concentrations of mannose (greater than 526.21 micrograms/ml) were demonstrated in hydrolysates of sera from 26 of 36 (72%) experimentally infected rats tested 1 or more days after onset of infection. Significant increases (greater than 453.49 micrograms/ml) were also seen in humans with localized (6/15), transient (2/2), and disseminated (5/7) candidiasis. The concentrations were highest in patients with candidemia occurring either transiently or as part of disseminated infection. The concentrations in patients with localized candidiasis were lower but were still significantly greater than control. These data suggest that changes in mannose concentration, as measured, may serve as markers of candidiasis. The amount of mannose present may be in part a function of the extent of the infection.
Molecular epidemiology of Vibrio cholerae in the U.S. Gulf Coast
Authors: Kaper J B , Bradford H B , Roberts N C , Falkow S
Published: January 2020

Enterotoxigenic strains of Vibrio cholerae O-1, biotype El Tor, isolated from a case of cholera in Texas in 1973, an outbreak of cholera in Louisiana in 1978, and Louisiana sewage samples in 1980 and 1981 were analyzed for their genetic similarities. Chromosomal DNA was isolated from each strain, digested with restriction endonuclease, and analyzed by the Southern blot technique. A radioactive probe consisting of Escherichia coli heat-labile enterotoxin DNA detected cholera toxin gene sequences in these strains and demonstrated that the toxin gene sequence, if not the entire chromosomal DNA, is identical in these strains and distinctly different from other strains of V. cholerae isolated throughout the world. In addition, two strains of enterotoxigenic V. cholerae non-O-1 isolated from clinical cases, were analyzed and found to possess cholera toxin genes which differed in the DNA sequence from the V. cholerae O-1 strains. We concluded that a single strain of enterotoxigenic V. cholerae O-1 is resident in the U.S. Gulf Coast and that a second reservoir of cholera toxin genes exists in V. cholerae non-O-1 strains in Louisiana.
Azlocillin, a ureido penicillin active against Pseudomonas aeruginosa: interpretive zone standards and quality control parameters for tests with 75-mu g disks
Authors: Barry A L , Jones R N , Thornsberry C , Gavan T L
Published: January 2020

A nine-laboratory coordinated study was performed to establish tentative control limits for 75-mu g azlocillin disks tested against the standard control strain of Pseudomonas aeruginosa (ATCC 27853). Control limits for individual tests were 24 to 30 mm (25 to 29 mm for means of five separate tests). To establish interpretive zone standards for 75-mu g azlocillin disks, three separate laboratories each tested 93 strains of P. aeruginosa or related species. Geometric mean minimal inhibitory concentrations (MICs) were plotted against arithmetic mean zone diameters. Regression analyses were performed with zone diameter as the independent variable and also with MIC as the independent variable. The following interpretive categories were recommended: resistant, less than or equal to 14 mm (MIC, greater than 128 mu g/ml); intermediate, 15 to 17 mm (MIC, 128 mu g/ml); and susceptible, greater than or equal to 18 mm (MIC, less than or equal to 64 mu g/ml).
Monoclonal Antibody to Streptococcal Group B Carbohydrate: Applications in Latex Agglutination and Immunoprecipitin Assays
Authors: Ruch Frank E. , Smith Lisette
Published: January 2020

Two monoclonal mouse antibodies with specificities for group B streptococcal capsular antigens were evaluated in assays for the identification of group B streptococci (GBS). One of these antibodies (A9) was shown to precipitate group B carbohydrate antigen in reactions with both purified group B antigen and antigen present in autoclave or enzyme extracts of GBS. A9 antibody was also specific for group B antigen in gel diffusion reactions with extracts of Lancefield group A, B, C, D, F, and G streptococci and was a highly sensitive reagent in detecting soluble group B antigen by counterimmunoelectrophoresis. Antigen extracted from all five serotypes of GBS was shown to be precipitated by A9 antibody. A second monoclonal antibody (C8) was reactive with intact GBS but did not precipitate soluble antigen in bacterial extracts. In contrast with what has been shown for polyclonal rabbit anti-group B antiserum, neither antibody was significantly inhibited in binding or precipitation assays by high concentrations of free rhamnose or other monosaccharides of carbohydrates found in group B antigen. Rhamnose, the most abundant carbohydrate of the group B antigen, does not appear therefore to be an immunodominant determinant in the binding of A9 or C8 antibody. The epitopes of both monoclonal antibodies are exposed on the surface of live as well as heat-fixed GBS cells. A9 antibody-coated latex particles were compared with a commercially available polyclonal latex agglutination reagent and shown to be equally sensitive and specific in the detection of soluble group B antigen in urine and cerebrospinal fluid from patients with GBS infections. Because of its uniformity and defined antigen specificity, which includes reactivity with all five serotypes of GBS, A9 antibody offers the potential of an improved immunodiagnostic reagent for the identification of GBS.
Rapid diagnosis of parainfluenza virus infection in children
Authors: Wong D T , Welliver R C , Riddlesberger K R , Sun M S , Ogra P L
Published: January 2020

The indirect immunofluorescent-antibody (IFA) assay for detection of parainfluenza virus antigen was performed on samples of nasopharyngeal secretions obtained from infants and children with various respiratory illnesses to determine the usefulness of the IFA assay for rapid diagnosis of parainfluenza virus infection. Ninety-four samples of nasopharyngeal secretions were obtained from 78 children during a community outbreak of parainfluenza virus infection. Application of the IFA assay revealed the presence of parainfluenza antigen in 67 of the 94 samples. When these same specimens were inoculated into tissue culture, the presence of parainfluenza virus was confirmed in 62 (93%) of the 67 IFA-positive specimens by hemadsorption on tissue culture monolayers (31 cases), presence of parainfluenza hemagglutinin in infected tissue culture fluids (23 cases), or by other methods (8 cases). Only four IFA-negative specimens were subsequently shown to be positive for parainfluenza virus by tissue culture infectivity. The IFA assay provided a more rapid and more accurate method for diagnosis of parainfluenza infection in children than did routine tissue culture methods employed currently.
Acridine orange staining of Pneumocystis carinii
Authors: Thomson R B , Smith T F
Published: January 2020

Acridine orange was used to stain smears of mouse lung which contained cyst and trophozoite forms of Pneumocystis carinii. Trophozoite forms stained yellow to orange; however, cyst forms did not stain. Acridine orange is a rapid and sensitive method for demonstrating trophozoites of P. carinii in mouse lung tissue.
Semiselective medium for isolation of Moraxella bovis from cattle with infectious keratoconjunctivitis
Authors: Webber J J , Fales W H , Selby L A
Published: January 2020

The incorporation of 2.5 micrograms/ml of cloxacillin into 5% bovine blood agar provided an inexpensive, easily prepared culture medium for the primary isolation of Moraxella bovis from bovine lacrimal and nasal secretions. With this medium, the time required to identify and isolate M. bovis from large numbers of field specimens was substantially reduced, whereas the sensitivity of isolation was increased by 60%.
Panophthalmitis caused by Vibrio parahaemolyticus
Authors: Tacket C O , Barrett T J , Sanders G E , Blake P A
Published: January 2020

We report a case of Vibrio parahaemolyticus panophthalmitis which resulted from contamination of a wound with water from a pond in inland Georgia. The pond was on the property of an oil refinery which receives crude oil from southern Mississippi. Cultures of the pond water 5 years later did not yield V. parahaemolyticus, but did yield non-O1 V. cholerae and had 0.28% sodium chloride content. V. parahaemolyticus may have been introduced into the pond along with oil transported from the Gulf of Mexico, and growth of this halophilic species may have been supported by salt from spilled crude oil.
Evaluation of a solid-phase immunofluorescence assay for detection of antibodies to Legionella pneumophila
Authors: Thompson T A , Wilkinson H W
Published: January 2020

A semiautomated solid-phase immunofluorescence technique (FIAX) was compared with the standard indirect immunofluorescence assay (IFA) for the determination of antibody levels to Legionella pneumophila serogroup 1 in paired human serum samples. The FIAX method was in agreement with the IFA test for 91.8% of the serum pairs but gave evidence of a recent Legionella infection significantly fewer times than did the IFA. These results suggest that the FIAX technique may eventually be a useful alternative test for measuring Legionella antibodies. However, further study will be required to determine its efficacy in providing a serodiagnosis of legionellosis.
Antigenic characterization of Cryptococcus neoformans serotypes and its application to serotyping of clinical isolates
Authors: Ikeda R , Shinoda T , Fukazawa Y , Kaufman L
Published: January 2020

Antigenic analysis of the four serotypes of Cryptococcus neoformans was carried out by slide agglutination with reciprocal adsorption methods. With this procedure the antigenic patterns of the serotypes were established. Serotypes A and D had antigenic factors 1, 2, 3, 7 and 1, 2, 3, 8, respectively. Serotypes B and C were found to have antigenic factors 1, 2, 4, 5 and 1, 4, 6, respectively. Factor sera, prepared according to the antigenic patterns demonstrated by adsorption studies, proved to be useful for rapidly and accurately identifying C. neoformans serotypes. Some patterns similar to those of the C. neoformans serotypes were observed in five other Cryptococcus species and two Candida species. The proton magnetic resonance spectra of polysaccharides from the C. neoformans serotypes correlated well with their antigenic characteristics. Phenol oxidase test reactions and growth at 37 degrees C were useful criteria for determining which yeasts should be chosen for clinical application of factor sera for serotyping of C. neoformans. Sixty-two Japanese isolates of C. neoformans were serotyped. Fifty-eight of these isolates were serotype A, three were serotype A-D, and one was serotype D.
Development of an enzyme-linked immunosorbent assay for studying Vibrio cholerae cell surface antigens
Authors: Cryz S J , Fürer E , Germanier R
Published: January 2020

An enzyme-linked immunosorbent assay for the measurement of antibodies directed against cell surface antigens of Vibrio cholerae (CSA ELISA) was developed. NaN3-killed whole cells of V. cholerae, adsorbed to polystyrene tubes, were used as immobilized antigens. The assay was capable of detecting antibodies directed against lipopolysaccharide and non-lipopolysaccharide surface antigens. In addition, the CSA ELISA was capable of detecting non-vibriocidal antibody. An antiserum raised in rabbits by immunization with live V. cholerae 1418 (Ogawa, El Tor) was capable of reacting with various heterologous strains of V. cholerae used as immobilized antigens. Therefore, common antigens shared by V. cholerae strains could be detected by using the CSA ELISA.
Characterization of nontypable Streptococcus pneumoniae-like organisms isolated from outbreaks of conjunctivitis
Authors: Shayegani M , Parsons L M , Gibbons W E , Campbell D
Published: January 2020

From January through June 1980 seven colleges and universities in various parts of New York State (NYS) reported outbreaks of conjunctivitis affecting at least 1,500 students of both sexes. Of the 125 conjunctival swabs tested in our laboratory, organisms identified as nontypable Streptococcus pneumoniae were isolated in pure culture from 24% and in combination with other organisms from 22%. Although bile-soluble and susceptible to optochin, the isolates had a dry-colony appearance and no typable capsule with the Neufeld capsular-swelling test. Mouse passage of four representative NYS isolates did not stimulate production of a typable capsule. We subsequently chose to refer to these isolates as S. pneumoniae-like organisms. Of primary importance to our study, all NYS isolates tested were similar in biochemical and immunological reactions, antibiotic susceptibility, and virulence in mice. Of 18 strains referred to us from three other outbreaks (California, 1980; NYS, 1981; Illinois, 1981), four of the six tested biochemically gave the same biochemical reactions as the four NYS isolates, and 16 of the 18 tested immunologically reacted strongly with antisera produced against those four isolates, showing line(s) of identity with each other and with the NYS isolates.
Comparison of direct and standard microtiter broth dilution susceptibility testing of blood culture isolates
Authors: Kiehn T E , Capitolo C , Armstrong D
Published: January 2020

Turbid broth (0.5 ml) from blood culture bottles was inoculated into 0.5 ml of brain heart infusion broth, incubated for 3 to 6 h, diluted 1:500 in distilled water, and then inoculated directly into microtiter broth dilution susceptibility trays to test for minimal inhibitory concentrations. The results were compared to the standard tests performed 24 h later on colonies from subculture plates. The minimal inhibitory concentrations measured by these two methods were compared in 1,875 organism-antibiotic tests. The two minimal inhibitory concentrations were identical in 86.0% and within one twofold dilution in 98.0% of the tests. An organism was judged to be susceptible by one method and resistant by the other in 13 tests (0.7%). These 13 discrepancies were distributed among several organism-antibiotic combinations; no more than two were seen for any one combination. Highly accurate susceptibility testing can be achieved by using direct inoculation of turbid blood culture broths.
In Vitro Studies with Cefotaxime: Disk Diffusion Susceptibility Tests
Authors: Shadomy Smith , Chan Edward L.
Published: January 2020

Until recently two sets of conflicting interpretive criteria existed for use with the standardized 30-μg cefotaxime diffusion disk [National Committee for Clinical Laboratory Standards (NCCLS) M2-A2S, supplement 1; product information package insert for Claforan (cefotaxime sodium), edition 10/81.] The latter criteria recently were superseded by a third set which differs radically from the first two in both minimal inhibitory concentration (MIC) and zone size breakpoints. The first two sets of criteria differed mainly in the zone of inhibition diameters used to predict resistant and intermediate organisms. The accuracies of these two sets of criteria were evaluated by paired agar dilution (World Health Organization-International Collaborative Study) and disk diffusion tests (Food and Drug Administration) with 347 clinical isolates of aerobic bacteria. A total of 274 isolates (79%) were clinically susceptible by agar dilution as defined by NCCLS (MIC ≤8 μg/ml) and 61, including 48
Pseudomonas
spp., were of intermediate susceptibility (MIC of 16 or 32 μg/ml). Twelve were resistant (MIC ≥64 μg/ml). The original product information package insert criteria proved to be most unreliable in identification of the intermediate organisms (zone of inhibition diameters of 18 to 22 mm); only 5 were correctly predicted as being intermediate, whereas 54 were predicted as being resistant, and 2 were predicted as being susceptible. In contrast, the NCCLS criteria (zone of inhibition diameter of 15 to 22 mm) predicted 41 as being intermediate and 18 as being resistant; again, 2 were susceptible. The 12 isolates resistant to cefotaxime by agar dilution were correctly predicted to be resistant by either set of breakpoints (zone of inhibition diameter of ≤14 or ≤17 mm). These results suggest that the package insert criteria previously recommended for use with the 30-μg cefotaxime disk were overly conservative and required revision to bring them into agreement with NCCLS performance standards. Unfortunately however, the recent revision of the package insert criteria has further confused this issue [product information package insert for Claforan (cefotaxime sodium), edition 2/82].
Clinical significance of coagulase-negative staphylococci
Authors: Sewell C M , Clarridge J E , Young E J , Guthrie R K
Published: January 2020

Although coagulase-negative staphylococci (C-NS) have been implicated in certain human infections, they are generally regarded as contaminants, and their clinical significance is questioned. To assess their role as pathogens, we studied 205 isolates of C-NS from wounds and body fluids (blood, urine, pleural and peritoneal fluids, etc.). Patient's charts were reviewed, and, by using strict criteria, a determination was made regarding the clinical significance of these isolates. The organisms were then identified to determine whether certain species of C-NS were associated with specific infections. S epidermidis sensu stricto accounted for 81% of the C-NS isolated. The frequencies of other species were: S. haemolyticus (6%), S. hominis (5%), S. capitis (4%), S. warneri (3%), and others (1%). Only two isolates were novobiocin resistant; neither was identified as S. saprophyticus. By using our criteria, 22% of the C-NS were considered to be clinically significant, and the majority of these (93%) was S. epidermidis. The most common source of the clinically relevant C-NS isolates was wounds. These data suggest that identification of C-NS species other than S. epidermidis may be of limited value in predicting clinical significance.
Collaborative evaluation of the micro-media systems anaerobe susceptibility panel: comparisons with reference methods and test reproducibility
Authors: Jones R N , Barry A L , Cotton J L , Sutter V L , Swenson J M
Published: January 2020

The Micro-Media Systems (MMS) anaerobe susceptibility testing panel results from four laboratories were compared for interlaboratory and intralaboratory variations and for the results with the reference agar dilution and a broth microdilution method. The interlaboratory agreement was 98.0% and intralaboratory agreement was 97.3% (+/- 1 log2 dilution). When interpretive criteria for each antimicrobial agent (susceptible, intermediate, and resistant) were assigned, the MMS anaerobic minimum inhibitory concentration data showed an interpretive accuracy of 91.0 and 95.5% for comparisons to the reference agar dilution and the broth methods, respectively. Most significant interpretive errors were considered minor, and nearly half of all errors involved tetracycline, a drug rarely used for serious anaerobic infections. The MMS anaerobe panels appear to be acceptable for selected use in clinical microbiology laboratories.
Hospital outbreaks caused by Pseudomonas aeruginosa: importance of serogroup O11
Authors: Farmer J J , Weinstein R A , Zierdt C H , Brokopp C D
Published: January 2020

Suspected outbreaks caused by pseudomonas aeruginosa in 19 hospitals and two motels were studied. On the basis of epidemiological analysis, serological typing, and antibiotic resistance patterns, 17 were classified as single-strain outbreaks. Six were classified as common-source outbreaks: of these, three were caused by contaminated urological instruments or solutions, two involved bathing in contaminated whirlpools, and one was caused by contaminated lens prostheses implanted during eye surgery. The ability of P. aeruginosa to survive or grow in wet environments was important in each of these six outbreaks. Eight outbreaks were classified as cross-infection. Two involved the urinary tract and were caused by antibiotic-resistant strains. Six involved the respiratory tract, but only one was caused by an antibiotic-resistant strain. In 2 of the 17 single-strain outbreaks, the exact mode of transmission could not be determined. One was an outbreak of pseudobacteremia in which patient blood cultures were contaminated with a single strain, presumably during collection of specimens or culture processing, P. aeruginosa serogroup O11 caused 9 of 17 (53%) single-strain outbreaks, a surprising finding since this serogroup represents only about 8% of endemic hospital isolates of this species. Serotyping was very useful in epidemiological analysis, but antibiotic susceptibility patterns were less useful.
Serotyping of Campylobacter jejuni isolated from sporadic cases and outbreaks in British Columbia
Authors: McMyne P M , Penner J L , Mathias R G , Black W A , Hennessy J N
Published: January 2020

Campylobacter jejuni from sporadic cases and outbreaks of gastroenteritis were serotyped on the basis of heat-extracted soluble thermostable antigens identified with the use of the passive hemagglutination technique. A total of 168 isolates were separated into 45 different types. The largest proportion of the isolates fell into three serotypes, each with 11 to 12.5% of the total number. Three less frequently occurring serotypes each included approximately 5%, and the remaining 50% of the isolates were distributed among 39 other serotypes. In most cases, serotyping demonstrated that epidemiologically linked isolates were of the same serotype, but the outbreak strains could belong either to frequently or to infrequently isolated serotypes. The high correlation between clinical findings and serotyping results confirmed the applicability of the serotyping scheme in epidemiological investigations of C. jejuni infections.
Detection of mixed Mycoplasma species
Authors: Bradbury J M , McClenaghan M
Published: January 2020

Immunofluorescence can be used to demonstrate mixed species within a mycoplasma culture; however, it may fail to do so if one species is present in very low numbers. To enhance the detection of minor components in such mixtures, a technique was developed, based on the growth inhibition test, whereby the growth of the major component in a mixed culture was inhibited, thus permitting the minor component to emerge. The method was applied to 67 field isolates from chickens and turkeys, which had been examined by immunofluorescence and were thought to contain only one mycoplasma species. Of these, 26 cultures were found to be mixtures, and in some instances the pathogenic species Mycoplasma gallisepticum and M. meleagridis were revealed.
Enzyme-linked immunosorbent assay for detection of respiratory syncytial virus infection: development and description
Authors: Hendry R M , McIntosh K
Published: January 2020

An indirect enzyme-linked immunosorbent assay for detection of respiratory syncytial virus (RSV) antigens was developed, using commercially available antisera. Horse anti-RSV and calf antiserum to bovine RSV were used as capture and detector antibodies, respectively. The assay could detect as few as 50 PFU of unpurified RSV per ml in infected cell culture supernatant fluids and as little as 10 ng of affinity-purified RSV antigen per ml. No cross-reactions were observed with heterologous virus types. Freeze-thaw treatment had no effect on RSV enzyme-linked immunosorbent assay titers, but viral transport medium inhibited RSV enzyme-linked immunosorbent assay titers from 10- to 100-fold. The assay can be easily performed in 24 h and is a sensitive and specific method for the detection of RSV antigens.
Chemiluminescent immunoenzymatic assay for rapid diagnosis of viral infections
Authors: Pronovost A D , Baumgarten A , Andiman W A
Published: January 2020

Current methods of viral diagnosis have been criticized for slowness and insensitivity. However, immunoassay may provide the desired increase in the speed of diagnosis without sacrificing accuracy. This report describes the efficacy of the detection of viral antigen by means of an absorptiometric enzyme-linked immunoassay (ELISA) or by a chemiluminescent enzyme-linked immunoassay (CELISA). Human cytomegalovirus was detected in clinical specimens or culture fluid with comparable sensitivity by CELISA and by viral isolation but with 50 times lesser sensitivity by ELISA. Similarly, herpes simplex virus was detected in clinical specimens with markedly greater sensitivity by CELISA than by ELISA. These findings indicate that the detection of appropriate viruses by CELISA may be a practical alternative to their isolation in cell culture.
Immunospecificity and quantitation of an enzyme-linked immunosorbent assay for group B streptococcal antibody
Authors: Anthony B F , Concepcion N F , McGeary S A , Ward J I , Heiner D C , Shapshak P , Insel R A
Published: January 2020

Type-specific antigen was purified from the supernatant of type III group B streptococcal cultures, tyrosylated, and bound to microtiter wells for an enzyme-linked immunosorbent assay. The immunological specificity of the antigen and the assay was shown by (i) reaction only with homologous unabsorbed rabbit sera and (ii) inhibition after incubation of human serum with homologous but not heterologous purified antigen. The assay was quantitated by relating optical density readings to absolute amounts of human immunoglobulin G bound to the microtiter wells.
Potassium tartrate-glycerol as a density gradient substrate for separation of small, round viruses from human feces
Authors: Ashley C R , Caul E O
Published: January 2020

Cesium chloride density gradients are frequently used for virus concentration or purification in the preparation of human feces for examination by electron microscopy, Disruption of some of the fecal viruses occurs if they are pelleted from the density gradient in an additional concentration step. This report highlights an important limitation imposed by the use of cesium chloride as a density gradient substrate in attempting to recover small, round, virus-like particles from feces and suggests an alternative substrate which preserves virus morphology without the use of additional protective agents.
Identification of clinical isolates of mycobacteria with gas-liquid chromatography: a 10-month follow-up study
Authors: Tisdall P A , DeYoung D R , Roberts G D , Anhalt J P
Published: January 2020

Identification of routine mycobacterial isolates by gas-liquid chromatography profile analysis was performed on 335 strains received at the Mayo Clinic over a 10-month period. Comparison of identification by gas-liquid chromatography versus conventional biochemical profiles was made. The two methods agreed on the identification of 320 isolates, with gas-liquid chromatography profiling making eight errors and biochemical profiling making four errors. In three cases, discrepancies could not be resolved.
Automated, quantitative cytopathic effect reduction assay for interferon
Authors: Yeh T J , McBride P T , Overall J C , Green J A
Published: January 2020

A rapid, cytopathic effect reduction assay for human interferon (IFN) is described. Dilutions of IFN were made with an automated diluter in 96-well microtiter plates. Total incubation time was 26 h. IFN titers were calculated from optical density readings of crystal violet-stained monolayers in an automated spectrophotometer, which required less than 1 min to read each plate.
Confirmation of aerogenic strains of Shigella boydii 13 and further study of Shigella serotypes by DNA relatedness
Authors: Brenner D J , Steigerwalt A G , Wathen H G , Gross R J , Rowe B
Published: January 2020

Shigella boydii 13 strains are separable from other Shigella and Escherichia coli strains on the basis of DNA relatedness. From this observation, it was possible to confirm the existence of aerogenic S. boydii 13 strains. DNA relatedness studies also showed that strains of E. coli and strains representing all other serotypes of Shigella, including provisional strains, belong to the same genetic species.
Sodium desoxycholate-extracted treponemal antigen in an enzyme-linked immunosorbent assay for syphilis
Authors: Hunter E F , Farshy C E , Liska S L , Cruce D D , Crawford J A , Feeley J C
Published: January 2020

The extraction of Treponema pallidum antigen with sodium desoxycholate, based on a previously described procedure (J. Portnoy and H.J. Magnuson, J. Immuno. 75:348-355, 1955), was used in an enzyme-linked immunosorbent assay (ELISA) test for syphilis. The antigen was prepared from T. pallidum street strain no. 14, and its overall sensitivity and specificity was compared with those of sonicated antigen preparations made with phosphate-buffered saline. The optimum serum dilution for testing and the significant absorbance reading at 490 nm were selected by examination of quantitative dilutions of 91 sera from presumably normal individuals and 92 sera from syphilitics. The time and temperature of serum and conjugate incubations were also examined. With an absorbance reading of greater than or equal to 0.2 at the 1:80 serum dilution, 88 (95.8%) of 92 sera from syphilitics were reactive in the ELISA test with desoxycholate-extracted antigen, and 82 (89.1%) were reactive with the sonicated antigen. Only one nonsyphilitic serum was reactive with each antigen. Greater sensitivity without loss in specificity was obtained with longer serum and conjugate incubations. We concluded that an ELISA test with sodium desoxycholate-extracted antigen is more sensitive than and equally specific to an ELISA with sonicated treponemal antigen.